Plants exhibiting reduced damage-induced surface discoloration

Abstract

To provide a plant showing reduced damage-induced surface discoloration.SOLUTION: The present invention relates to a plant including modified-type F5H gene homolog, where the gene homolog includes modification comparing to its corresponding wild-type F5H gene homolog, and the presence of the modified-type F5H gene homolog in a plant brings reduction of damage-induced surface discoloration comparing to a plant that does not include the modified-type F5H gene homolog. The present invention also relates to a modified-type F5H gene homolog bringing reduced damage-induced surface discoloration. The present invention further relates to use of the gene in breeding and production in a plant showing reduced damage-induced surface discoloration.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to plants that exhibit reduced wound-induced surface discoloration. The present invention further relates to modified gene homologs that result in reduced wound-induced surface discoloration in plants, their sequences, and the use of these sequences to identify the presence of the modified gene homologs. The present invention also relates to seeds and progeny of such plants, as well as seedlings for obtaining such plants. [Background technology]

[0002] In recent years, consumer convenience and the development of new varieties have contributed to an expansion of the selection of commercially available processed vegetables and fruits. Ready-to-eat products, i.e., products in pre-cut, pre-washed, and packaged forms, may contain lettuce (Lactuca sativa) and other leafy vegetables, such as chicory (Cichorium intybus) and endive (Cichorium endivia), as individually processed or mixed products. One of the most important and frequently encountered problems during the harvesting, processing, and storage of vegetables is the occurrence of wound-induced surface changes, which are visible discoloration due to pink discoloration of the surface of the plant or its parts, gradually changing to brown after long-term storage. Other crops, such as potato (Solanum tuberosum), onion (Allium cepa), artichoke (Cynara cardunculus var. Scolymus), rice (Oryza sativa), corn (Zea mays), peach (Prunus persica), eggplant (Solanum melongen), celery (Apium graveolens), apple (Malus domestica), banana (Musa acuminate), soybean (Glycine max), pear (Pyrus x bretschneideri), wheat (Triticum aestivum), radish (Raphanus sativus), cabbage, and cauliflower (Brassica oleracea), may also suffer from visible damage-induced surface discoloration of the plant or parts thereof, such as leaves, whole plant heads, fruits, inflorescences, seeds, endosperm, stems, tubers, bulbs, and roots.

[0003] Wound-induced surface discolouration, or wound-induced discolouration, is caused by a strong wound response at and around the wound site, resulting in rapid discoloration of the harvested and optionally processed produce, which consumers consider to be unattractive and potentially impair the quality of the produce, leading to a reduction in the market value of the produce and / or the disposal of the harvested and optionally processed produce.

[0004] The wound response is the response of a plant or its parts to heal a wound and protect itself from pathogens by creating a new insulation barrier. It is a complex biological response of plants to physical damage, such as cutting or bruising, and involves the activity of numerous proteins. The local response is primarily aimed at closing the wound surface, which is achieved by localized cell death at or immediately behind the wound surface. In addition to these visible effects, other responses, such as increased respiration or increased ethylene production, are known to be induced.

[0005] Studies at the biochemical level have shown that wounding can induce, inter alia, the phenylpropanoid pathway (PP pathway), which is required for the production of polyphenols and other compounds important to plants.

[0006] The first step in the PP pathway is the conversion of the amino acid phenylalanine to cinnamic acid by phenylalanine ammonia-lyase (PAL). PAL is enhanced upon wound injury by the induction of gene expression of at least one of its isoforms. This response results in the formation of polyphenols that are oxidized by polyphenol oxidase (PPO). PPO is present in plastids and is released and activated upon wounding. Oxidation of polyphenols leads to the formation of highly reactive quinones, which react with amino acids or proteins to produce pink, brown, or black discoloration.

[0007] Many post-harvest and post-processing treatments have been developed and applied to reduce damage-induced surface discolouration of vegetables such as lettuce. Examples of chemical or physical treatments are modified atmosphere packaging of fresh-cut leafy vegetables, application of edible coatings, heat shock treatments and application of chemicals.

[0008] Although these treatments prevent the appearance of wound-induced discoloration, the harvested and ultimately processed produce is still susceptible to discoloration if the packaging is damaged or if damage occurs shortly after the packaging is opened. Furthermore, the use of chemicals and the need for specialized equipment for such treatments significantly increase costs. For these reasons, more feasible genetically based solutions that act to reduce wound-induced surface discoloration in plants are preferred. Summary of the Invention

[0009] In the research leading to the present invention, it was surprisingly found that modifying F5H gene homologs in plants reduces wound-induced surface discoloration compared to plants or parts thereof that do not contain such modifications in their corresponding wild-type F5H gene homologs. F5H gene homologs encode ferulate 5-hydroxylase (F5H) protein homologs. Two F5H gene homologs have been described in Arabidopsis thaliana, designated F5H1 and F5H2. The F5H enzyme is part of the PP pathway, which is responsible for the hydroxylation of coniferyl aldehyde and coniferyl alcohol. The F5H protein belongs to a novel family of plant cytochrome P450-dependent monooxygenases called CYP84. However, the involvement of F5H in wound-induced surface discoloration has not yet been described.

[0010] It is an object of the present invention to provide plants that exhibit reduced wound-induced surface discolouration.

[0011] Thus, the present invention provides a plant comprising a modified F5H gene homolog, wherein the gene homolog comprises a modification relative to its corresponding wild-type F5H gene homolog, and wherein the presence of the modified F5H gene homolog in the plant results in reduced wound-induced surface discoloration compared to a plant that does not contain the modified F5H gene homolog.

[0012] The term "reducing" is usually measured relative to the wound-induced surface discoloration of a control plant or portion thereof that does not have such an alteration in its F5H gene homolog and thus does not exhibit reduced wound-induced surface discoloration, even though the control plant or portion thereof is a wild-type plant containing a wild-type F5H gene homolog. As used herein, a plant exhibiting "reduced wound-induced surface discoloration" or "reduced wound-induced surface discoloration" is a plant that has reduced wound-induced surface discoloration compared to the wound-induced surface discoloration of a wild-type plant. Thus, an improvement in reduced wound-induced surface discoloration is defined by a delayed appearance and / or reduced intensity of the discoloration compared to a plant that does not contain a modified F5H gene homolog. The reduced intensity of the discoloration is visible as a lower discoloration intensity and / or a smaller discolored surface of the wounded surface compared to the discoloration and surface of the wound-induced surface discoloration of a wild-type plant. Ultimately, wound-induced discoloration is completely absent. A delayed appearance of discoloration means that the onset of discoloration is delayed. Thus, the plant maintains its fresh appearance longer, which in effect results in an extended shelf life.

[0013] The present invention further provides sequences of modified F5H gene homologs for identifying plants containing modifications that lead to the traits of the present invention.

[0014] The number of F5H gene homologs within a particular species varies among different plant species. According to the definition of gene homologs described herein, the lettuce (Lactuca sativa) plant genome was found to contain two F5H gene homologs. One homolog, designated herein as F5H1, is located on chromosome 4, has a wild-type DNA coding sequence (CDS) set forth in SEQ ID NO: 115, and encodes the wild-type F5H1 protein of SEQ ID NO: 1. The other lettuce homolog, designated herein as F5H2, is located on chromosome 3, has a wild-type DNA coding sequence set forth in SEQ ID NO: 116, and encodes the wild-type F5H2 protein of SEQ ID NO: 2.

[0015] The genomes of artichoke (Cynara cardunculus var. Scolymus), rice (Oryza sativa), maize (Zea mays), peach (Prunus persica), and eggplant (Solanum melongen) were also found to contain two wild-type F5H gene homologs with SEQ ID NOs listed in Table 2. The plant species chicory (Cichorium intybus), endive (Cichorium endivia), celery (Apium graveolens), and apple (Malus domestica) contain three wild-type F5H gene homologs with SEQ ID NOs listed in Table 2. The genome of banana (Musa acuminate) contains four wild-type F5H homologs with SEQ ID NOs listed in Table 2. The genomes of soybean (Glycine max), pear (Pyrus x bretschneideri), wheat (Triticum aestivum), radish (Raphanus sativus), and cabbage / cauliflower (Brassica oleracea) contain five wild-type F5H homologs with the SEQ ID NOs listed in Table 2, while the genomes of potato (Solanum tuberosum) and onion (Allium cepa) contain one wild-type F5H gene homolog in their genomes with the SEQ ID NOs listed in Table 2.

[0016] In a preferred embodiment, the present invention relates to a plant belonging to the genus Lactuca. In particular, the present invention relates to a lettuce plant comprising two F5H gene homologs that are modified compared to the nucleotide sequence of the wild-type gene (SEQ ID NOs: 115 and 116), which encode the wild-type proteins (SEQ ID NOs: 1 and 2). Other plants containing modified F5H gene homologs in their genomes, such as potato (Solanum tuberosum), onion (Allium cepa), artichoke (Cynara cardunculus var. Scolymus), rice (Oryza sativa), corn (Zea mays), peach (Prunus persica), eggplant (Solanum melongen), chicory (Cichorium intybus), endive (Cichorium endivia), celery (Apium graveolens), apple (Malus domestica), banana (Musa acuminate), soybean (Glycine max), pear (Pyrus x bretschneideri), wheat (Triticum aestivum), radish (Raphanus sativus), and cabbage / cauliflower (Brassica oleracea), are also part of the present invention.

[0017] In another aspect, the present invention relates to a modified F5H gene homolog in a plant belonging to the plant species potato, onion, lettuce, artichoke, rice, maize, peach, eggplant, chicory, endive, celery, apple, banana, soybean, pear, wheat, radish, or cabbage / cauliflower, wherein the modified F5H gene homolog comprises at least one modification compared to its wild-type sequence, which modification confers reduced wound-induced surface discoloration in the plant. The modified F5H gene of Arabidopsis thaliana, or the sequence in the NCBI database under accession number XP_011028697 (predicted: cytochrome P450 84A1-like [Populus euphratica]) herein, is not intended to be encompassed by the present invention.

[0018] In one aspect, the present invention relates to a modified F5H1 gene homolog having the sequence set forth in SEQ ID NO: 174, which confers reduced wound-induced surface discolouration compared to a plant that does not contain the modified F5H1 gene homolog.

[0019] In one aspect, the invention relates to a modified F5H1 gene homolog having the sequence set forth in SEQ ID NO: 174 and a modified F5H2 gene homolog having the sequence set forth in SEQ ID NO: 175, 176, 177, 178 or 179, which result in reduced wound-induced surface discoloration compared to a plant that does not contain the modified F5H gene homolog. The invention relates to a plant comprising a modified F5H gene homolog, wherein the gene homolog comprises a modification compared to its corresponding wild-type F5H gene homolog, and wherein the presence of the modified F5H gene homolog in the plant results in reduced wound-induced surface discoloration compared to a plant that does not contain the modified F5H gene homolog.

[0020] In one aspect, the present invention relates to a plant comprising a modified F5H gene homolog, wherein the gene homolog comprises a modification relative to its corresponding wild-type F5H gene homolog, and wherein the presence of the modified F5H gene homolog in the plant results in reduced wound-induced surface discoloration compared to a plant that does not contain the modified F5H gene homolog, and wherein the wild-type F5H gene sequence is set forth in any one of SEQ ID NOs: 58 to 114.

[0021] In a particular embodiment, the present invention relates to a plant comprising a modified F5H gene homolog of the present invention, wherein the plant is selected from the group consisting of potato, onion, lettuce, artichoke, rice, corn, peach, eggplant, chicory, endive, celery, apple, banana, soybean, pear, wheat, radish, and cabbage / cauliflower, and whose wild-type F5H gene sequence is the sequence of the SEQ ID NO: set forth in Table 2. As used herein, Arabidopsis plants comprising a modified F5H gene, in particular plants described in Meyer et al., National Academy of Sciences (1996) or Huang et al., Planta; an international journal of plant biology (2009), or plants comprising the sequence set forth in the NCBI database under accession number XP_011028697 (predicted: cytochrome P450 84A1-like [Populus euphratica]), are not intended to be encompassed by the present invention.

[0022] In another aspect, the present invention relates to a plant comprising two modified F5H gene homologs, wherein the presence of the modified F5H gene homologs in the plant results in reduced wound-induced surface discoloration compared to a plant that does not contain the modified F5H gene homologs.

[0023] In a particular embodiment, the present invention relates to a plant comprising two modified F5H gene homologs of the present invention, wherein the plant is selected from the group consisting of lettuce, artichoke, rice (Japonica), maize, peach, eggplant, chicory, endive, celery, apple, banana, soybean, pear, wheat, radish, and cabbage / cauliflower, and whose wild-type F5H gene sequence is the sequence of the SEQ ID NO: set forth in Table 2.

[0024] In another aspect, the present invention relates to a plant comprising three modified F5H gene homologs, wherein the presence of the modified F5H gene homologs in the plant results in reduced wound-induced surface discoloration compared to a plant that does not contain the modified F5H gene homologs.

[0025] In a particular embodiment, the present invention relates to a plant comprising three modified F5H gene homologs of the present invention, wherein the plant is selected from the group consisting of chicory, endive, celery, apple, banana, soybean, pear, wheat, radish, and cabbage, and whose wild-type F5H gene sequence is the sequence of the SEQ ID NO: set forth in Table 2.

[0026] In another aspect, the present invention relates to a plant comprising four modified F5H gene homologs, wherein the presence of the modified F5H gene homologs in the plant results in reduced wound-induced surface discoloration compared to a plant that does not contain the modified F5H gene homologs.

[0027] In a particular embodiment, the present invention relates to plants comprising four modified F5H gene homologs of the present invention, wherein the plants are selected from the group consisting of banana, soybean, pear, wheat, radish and cabbage / cauliflower, and whose wild-type F5H gene sequences are those of the SEQ ID NOs set forth in Table 2.

[0028] In another aspect, the present invention relates to a plant comprising five or more modified F5H gene homologs, wherein the presence of the modified F5H gene homologs in the plant results in reduced wound-induced surface discoloration compared to a plant that does not contain the modified F5H gene homologs.

[0029] In a particular embodiment, the present invention relates to a plant comprising five or more modified F5H gene homologs of the present invention, wherein the plant is selected from the group consisting of soybean, pear, wheat, radish and cabbage / cauliflower, and whose wild-type F5H gene sequence is the sequence of a SEQ ID NO: set forth in Table 2.

[0030] The present invention relates to edible plants, such as vegetables, fruits and crops, comprising a modified F5H gene homolog, wherein the gene homolog comprises a modification relative to its corresponding wild-type F5H gene homolog, and wherein the presence of the modified F5H gene homolog in the plant results in reduced wound-induced surface discoloration compared to a plant that does not contain the modified F5H gene homolog.

[0031] As used herein, a "plant of the invention" refers to a plant containing a modified F5H gene homolog, wherein the gene homolog contains a modification relative to its corresponding wild-type F5H gene homolog, and the presence of the modified F5H gene homolog in the plant results in reduced wound-induced surface discoloration compared to a plant that does not contain the modified F5H gene homolog. Plants of the invention are edible plants such as vegetables, fruits, and crops. Preferably, the plant of the invention is selected from the group consisting of potato, onion, lettuce, artichoke, rice, corn, peach, eggplant, chicory, endive, celery, apple, banana, soybean, pear, wheat, radish, and cabbage / cauliflower, and the wild-type F5H gene sequence thereof is a sequence of a SEQ ID NO: set forth in Table 2. Even more preferably, the plant of the invention is lettuce (Lactuca sativa).

[0032] The relationship between genes is defined as homologous (homozygous). As used herein, "homologous genes" refers to two related genes derived from a common ancestral gene. Homologous sequences are referred to as "homologs," and this term can apply to both genes and proteins. The terms "homologous" or "homolog" can be used interchangeably. Homologous genes encode homologous proteins. By our definition, the wild-type sequences of F5H protein homologs relevant to the present invention are set forth in Table 2. Furthermore, all F5H protein homologs identified during the research leading to the present invention share the five consensus sequence motifs shown in Table 1 and provided by MAST (Motif Alignment & Search Tool) and MEME (Multiple Em for Motif Elicitation) as sequences with any degree of variation in their amino acid sequences.

[0033] Preferably, an F5H protein homologue of the invention comprises a motif set forth in Table 1 with at least 54% identity, more preferably at least 60% identity, even more preferably at least 65% identity, even more preferably at least 70% identity, even more preferably at least 75% identity, even more preferably at least 80% identity, even more preferably at least 85% identity, even more preferably at least 90% identity, even more preferably at least 95% identity, and most preferably 100% identity. The consensus sequences for the five motifs are set forth in Table 1 and are highlighted in the alignment of F5H protein orthologues in Figure 4.

[0034] [Table 1]

[0035] "Orthologous genes" are homologous genes present in different species that originated from a common ancestral gene and diverged due to a speciation event. The terms "orthologous genes" or "orthologs" may be used interchangeably. Thus, the present invention provides modifications to F5H gene homologs within a species and to F5H gene orthologs in different species, all of which result in reduced wound-induced surface discoloration in the species. The wild-type sequences of F5H proteins and the sequence numbers of gene orthologs relevant to the present invention are set forth in Table 2.

[0036] As used herein, "gene" includes exon sequences and regulatory sequences such as promoter sequences, UTRs, and polyadenylation signals, and, when present, intron sequences. Modifications to an F5H gene homolog create a "modified gene homolog" by introducing at least one mutation in the nucleotide sequence of the gene. The terms "modification" and "mutation" may be used interchangeably. Generally, modifications alter gene expression and / or the activity of the protein encoded by the gene containing the modification. Modifications to the gene sequence may inhibit gene transcription such that expression of the modified gene is prevented or reduced, or may result in unstable mRNA. Modifications may also be changes to the sequence of the F5H gene that result in reduced activity levels, reduced activity, or a complete lack of activity of the encoded protein. In some cases, modifications may lead to overexpression of the protein, which may be responsible for an altered phenotype. Non-limiting examples of modification methods and techniques for modifying genes are described herein.

[0037] As used herein, "wild-type" or "WT" refers to the form of the organism (plant) as it occurs in nature, in the case of plants that do not exhibit reduced wound-induced surface discoloration.

[0038] As used herein, wild-type gene or gene homolog refers to an unmodified F5H gene, such as may result in a plant that does not exhibit reduced wound-induced surface discoloration. Wild-type plants are used as control plants that do not carry the modified F5H gene homolog and therefore do not exhibit reduced wound-induced surface discoloration. For comparability, plants of the present invention containing the modified F5H gene homolog and wild-type plants should be selected from the same species (type), preferably the same variety, of the same age, and grown under the same conditions.

[0039] As used herein, the term "F5H gene" or "modified F5H gene" refers to one or more modified F5H genes. A "plant comprising a modified F5H gene homolog" comprises one or more modified F5H gene homologs.

[0040] As used herein, the term "trait" refers to a plant phenotype. "Trait of the invention," "trait," or "phenotypic trait," "phenotype," and "characteristic" may be used interchangeably. As used herein, a trait of the invention is reduced wound-induced surface discoloration as a result of the presence of a modified F5H gene homolog and its corresponding F5H protein.

[0041] As used herein, "modified F5H gene homolog of the present invention" refers to an F5H homolog comprising any modification that results in reduced wound-induced surface discoloration in plants. Preferably, modified F5H gene homolog of the present invention comprises a modification shown in Table 3. "Modified F5H gene homolog of the present invention," "gene of the present invention," "F5H gene of the present invention," and "F5H gene homolog of the present invention" may be used interchangeably.

[0042] The present invention relates to a plant of the present invention, wherein the reduced endogenous level of F5H1 protein is due to a premature stop codon in the wild-type F5H1 sequence as set out in Table 2.

[0043] The present invention relates to a method for producing a plant that exhibits reduced wound-induced surface discolouration, the method comprising reducing the endogenous level of F5H1 protein in the plant.

[0044] The present invention further relates to a method of producing a plant that exhibits reduced wound-induced surface discolouration, comprising reducing the endogenous level of F5H1 protein in the plant, wherein the mutation is introduced by CRISPR, chemicals, radiation, or a combination thereof.

[0045] Modifications of the F5H gene can be introduced by mutagenesis. Several chemical or physical treatments are known to those skilled in the art and can be used to induce genetic mutations in plant species such as lettuce. Mutagenesis includes the random introduction of at least one modification by using one or more compounds such as ethyl methanesulfonate (EMS), nitrosomethylurea, hydroxylamine, proflavine, N-methyl-N-nitrosoguanidine, N-ethyl-N-nitrosourea, N-methyl-N-nitro-nitrosoguanidine, diethyl sulfate, ethyleneimine, sodium azide, formalin, urethane, phenol, and ethylene oxide, and / or physical means such as UV irradiation, fast neutron irradiation, X-rays, and gamma radiation, and / or by inserting genetic elements such as transposons, T-DNA, and retroviral elements. Mutagenesis also includes more specific targeted introduction of at least one modification by homologous recombination, oligonucleotide-based mutagenesis, zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs) or clustered regularly interspaced short palindromic repeats (CRISPR) systems.

[0046] Seeds of plants to be modified may be treated with solutions containing different concentrations of mutagens, such as EMS. EMS primarily alkylates guanine (G) residues in DNA strands, resulting in their pairing with thymine (T) instead of cytosine (C) during DNA replication. Thus, GC base pairs are converted to AT base pairs at a frequency determined by the effective dose of EMS and the activity of the plant's mismatch repair system. The effective dose of EMS varies depending on the concentration used, the size and other physical characteristics of the seeds, and the incubation time of the seeds in the EMS solution. Seeds treated with EMS are generally called M1. As a result of the treatment, M1 tissue contains random point mutations in the genome within its cells, and these are present in a subpopulation of cells that will form germline tissue (germocytes), which are inherited by the next generation, called M2. Mutations that are haploinsufficient and therefore result in sterility or embryonic lethality, or a combination thereof, are not inherited by the M2 generation. It should be noted that although most EMS induce mutations and the resulting traits are recessive (recessive), dominant (dominant) mutations leading to semi-dominant (semi-dominant) or dominant (dominant) traits can occur.

[0047] In one aspect, the present invention relates to a plant comprising a modified F5H gene homolog, wherein the gene homolog is mutated relative to its corresponding wild-type F5H gene homolog, and wherein the presence of the modified F5H gene homolog in the plant results in reduced wound-induced surface discoloration compared to a plant that does not contain the modified F5H gene homolog.

[0048] In certain aspects, the present invention relates to plants comprising a modified F5H gene homolog, wherein the gene homolog comprises an artificial mutation relative to its corresponding wild-type F5H gene homolog, and wherein the presence of the modified F5H gene homolog in the plant results in reduced wound-induced surface discoloration compared to plants that do not contain the modified F5H gene homolog.

[0049] In one aspect, the present invention relates to a plant or part of a plant comprising a modified F5H1 gene homolog of the present invention, wherein the modification results in reduced or absent protein expression of the F5H1 protein homolog compared to the expression of the protein produced by the corresponding wild-type F5H1 gene homolog.

[0050] In a particular embodiment, the present invention relates to a lettuce plant or part of a plant comprising a modified F5H1 gene homolog, wherein the modification results in reduced or absent protein expression of the F5H1 protein homolog compared to the expression of the protein produced by the corresponding wild-type F5H1 gene homolog.

[0051] In one aspect, the present invention relates to a plant or part of a plant comprising a modified F5H2 gene homolog of the present invention, wherein the modification results in a reduction, deletion or increase in protein expression of the F5H2 protein homolog compared to the expression of the protein produced by the corresponding wild-type F5H2 gene homolog.

[0052] In a particular embodiment, the present invention relates to a lettuce plant or part of a plant comprising a modified F5H2 gene homolog, wherein the modification results in a reduction, deletion or increase in protein expression of the F5H2 protein homolog compared to the expression of the protein produced by the corresponding wild-type F5H2 gene homolog.

[0053] The present invention further relates to a method for producing a plant of the present invention, comprising reducing the endogenous level of F5H1 protein in the plant, wherein reducing the endogenous level of F5H1 protein in the plant is achieved by reducing the expression of an F5H1 gene homolog in the plant by gene silencing or RNAi.

[0054] The present invention also relates to plants that exhibit reduced wound-induced surface discolouration and reduced F5H1 expression, wherein the reduction or loss is caused by the methods described herein.

[0055] When the expression of the modified F5H gene is deleted or reduced in the present invention, this means that gene expression leading to the synthesis of a functional protein is prevented, and therefore the expression of the modified F5H gene is deleted or is lower than that of the wild-type F5H gene, resulting in a low protein level. The prevention or reduction of gene expression is directly or indirectly responsible for the trait of reduced damage-induced surface discoloration herein.

[0056] Gene expression can also be inhibited or reduced by blocking the transcription of the gene, for example, using RNA or DNA oligonucleotides, or preferably by expressing a transcription factor that negatively affects the F5H gene promoter. Another example of a method for inhibiting or reducing gene expression is destabilizing F5H mRNA or transcripts by a nucleic acid molecule complementary to F5H mRNA or transcripts, preferably selected from the group consisting of antisense RNA, RNAi molecules, virus-induced gene silencing (VIGS) molecules, co-suppressor molecules, RNA oligonucleotides, or DNA oligonucleotides. Such methods for destabilizing mRNA or transcripts are well known to those skilled in the art.

[0057] Examples of modifications that result in reduced or absent F5H activity include modifications that lead to premature stop codons, frameshifts, or amino acid substitutions in the encoded protein. The reduced or absent F5H protein activity is directly or indirectly responsible for the reduced damage-induced surface discoloration trait herein. The reduced activity of the F5H protein can be achieved, for example, by introducing one or more mutations into the coding sequence of the F5H gene. The mutation(s) in the F5H gene can affect the biological function of the encoded protein compared to the F5H protein encoded by a wild-type F5H gene that does not contain such mutations.

[0058] In one aspect, the present invention relates to a plant exhibiting reduced wound-induced surface discolouration comprising a modified F5H gene homolog, wherein the modification results in a reduction or absence of protein activity of the F5H protein homolog compared to the activity of the protein produced by the corresponding wild-type F5H gene homolog.

[0059] In one particular embodiment, the present invention relates to a plant exhibiting reduced wound-induced surface discoloration comprising a modified F5H1 gene homolog, wherein the modification results in a reduction or absence of protein activity of the F5H1 protein homolog compared to the activity of the protein produced by the corresponding wild-type F5H1 gene homolog.

[0060] In one aspect, the present invention relates to a plant exhibiting reduced wound-induced surface discoloration comprising a modified F5H2 gene homolog, wherein the modification results in a reduction or absence of protein activity of the F5H2 protein homolog compared to the activity of the protein produced by the corresponding wild-type F5H2 gene homolog.

[0061] Plants of the present invention were generated using the mutagen EMS as described in Example 1. Plants grown from seeds treated once with the mutagen and selected for their ability to exhibit reduced wound-induced surface discoloration contain at least one mutation in one F5H gene homolog in their genome. Seeds already carrying one or more modified F5H gene homologs were treated several more times with the mutagen to introduce modifications into other F5H gene homologs in the plant, and selected for their ability to exhibit reduced wound-induced surface discoloration after each treatment with the mutagen. For example, a lettuce plant with two F5H gene homologs in its genome was treated twice with the mutagen EMS to introduce modifications into the plant's two F5H gene homologs.

[0062] Genetic modifications can be recessive, dominant, or intermediate. The terms "intermediate" and "semi-dominant" can be used interchangeably. In the case of recessive traits, the genetic modification must be present in the homozygous state for the trait to be fully expressed. Some of the modifications described herein are recessive and therefore confer reduced wound-induced surface discoloration only when both alleles of the gene have the modification. Modifications that are dominant or intermediate can also be expressed in the heterozygous state. The phenotype of a heterozygote for an intermediate trait is between that of a homozygous dominant genotype and a homozygous recessive genotype. These types of modifications are also part of the present invention.

[0063] The modification in the F5H1 or F5H2 gene homologue can be present in a heterozygous state or a homozygous state.Preferably, the modification in the F5H1 gene homologue is present in a homozygous state.The genotype of the plant can be confirmed using molecular markers.Preferably, the genotype of the plant can be confirmed using molecular markers as described in Example 3.

[0064] In one aspect, the present invention relates to plants comprising an F5H gene homolog that comprises an alteration relative to the corresponding wild-type sequence, wherein the alteration results in a distinct phenotype that exhibits reduced wound-induced surface discoloration.

[0065] In one aspect, the present invention relates to a plant comprising an F5H gene homolog that comprises a modification relative to the corresponding wild-type sequence, wherein the modification results in a distinct phenotype that exhibits reduced wound-induced surface discoloration compared to a plant that does not comprise the modified F5H gene homolog.

[0066] Herein, reduced wound-induced discolouration is preferably achieved by modifications present in the genome of lettuce seeds deposited with NCIMB under accession numbers NCIMB42546, NCIMB2547, NCIMB42548, NCIMB42549, NCIMB42550 and NCIMB42551. The modifications are set out in Table 3.

[0067] Seeds of seed lot 16E.607_B01 were deposited with NCIMB under accession number NCIMB 45546. The deposited seeds contain mutation 1 in the F5H1 gene homolog.

[0068] Seeds of seed lot 16E.607_B02 have been deposited with NCIMB under accession number NCIMB 42547. The deposited seeds contain mutation 1 in the F5H1 gene homolog and mutation 2 in the F5H2 gene homolog.

[0069] Seeds of seed lot 16E.607_B03 have been deposited with NCIMB under accession number NCIMB 42548. The deposited seeds contain mutation 1 in the F5H1 gene homolog and mutation 3 in the F5H2 gene homolog.

[0070] Seeds of seed lot 16E.607_B04 were deposited with NCIMB under accession number NCIMB 42549. The deposited seeds contain mutation 1 in the F5H1 gene homolog and mutation 4 in the F5H2 gene homolog.

[0071] Seeds of seed lot 16E.607_B05 were deposited with NCIMB under accession number NCIMB 42550. The deposited seeds contain mutation 1 in the F5H1 gene homolog and mutation 5 in the F5H2 gene homolog.

[0072] Seeds of seed lot 16E.607_B06 were deposited with NCIMB under accession number NCIMB 42551. The deposited seeds contain mutation 1 in the F5H1 gene homolog and mutation 6 in the F5H2 gene homolog.

[0073] The lettuce F5H1 gene homolog containing mutation 1 is shown in SEQ ID NO: 174.

[0074] The lettuce F5H2 gene homolog containing mutation 2 is shown in SEQ ID NO: 175.

[0075] The lettuce F5H2 gene homolog containing mutation 3 is shown in SEQ ID NO:176.

[0076] The lettuce F5H2 gene homolog containing mutation 4 is shown in SEQ ID NO: 177.

[0077] The lettuce F5H2 gene homolog containing mutation 5 is shown in SEQ ID NO: 178.

[0078] The lettuce F5H2 gene homolog containing mutation 6 is shown in SEQ ID NO: 179.

[0079] Modifications to a gene can result in premature stop codons, frameshifts, amino acid substitutions, or splice variants in the corresponding protein sequence. Modifications in protein sequences are the result of base pair substitutions, deletions, and changes in the DNA sequence encoding the protein.

[0080] When a modification to a DNA sequence results in a premature stop codon, its transcription results in a truncated form of the encoded protein. Modifications can occur in regions of the protein sequence that contain one or more domains or active sites essential for it to perform its function, and / or to interact with its substrates or other proteins, and / or to fold into a functional protein.

[0081] When modifications to a DNA sequence result in a frameshift mutation, protein translation often results in an amino acid sequence that is completely different from the wild-type sequence and a premature stop codon. The translated protein usually has a different biological function than the wild-type protein. These modifications involve the insertion or deletion of multiple base pairs that are not a multiple of three, resulting in a shift in the triplet codons encoding the protein's individual amino acids and therefore changing the protein's amino acid sequence compared to the original open reading frame. Insertions or deletions that are a multiple of three can also result in an amino acid sequence that differs from the wild-type sequence.

[0082] Alterations of one or more base pairs in the coding sequence of a DNA sequence can result in an amino acid change in the encoded protein sequence. Due to the redundancy of the genetic code, some mutations result in the same amino acid; these mutations are called "silent mutations." Furthermore, some amino acid changes are "conservative," i.e., they result in the substitution of one amino acid for another with similar properties, such as similar polarity, charge, solubility, hydrophobicity, hydrophilicity, or amphipathicity of the residue, so that the mutation does not dramatically affect the function of the mature protein and / or alter its folding. Other amino acid changes are non-silent, non-conservative amino acid changes, such as the substitution of one amino acid for another with different chemical properties in domains affecting substrate recognition, the active site of an enzyme, interaction domains, or key structural domains. Such amino acid changes can partially or completely disrupt the function of the encoded protein without affecting the expression level of the encoded gene. These types of mutations can have adverse effects on the stability, functionality, and / or structure of the encoded protein. Non-silent and non-conservative amino acid changes can also result in overexpression of the encoded protein.

[0083] Mutations in the promoter sequence of the F5H gene can also destabilize the biological function of the encoded F5H protein; for example, such mutations can result in a complete lack of transcription of the gene (e.g., resulting in a complete absence of the subsequent F5H protein), or in a significantly reduced or biologically inappropriate level of transcription (e.g., resulting in a reduced level of the subsequent F5H protein), or in overexpression of the F5H protein (e.g., resulting in a higher level of the subsequent F5H protein).

[0084] In the present invention, gene expression analysis was performed by measuring RNA of F5H1 gene homologs and F5H2 gene homologs. The analysis showed that expression of F5H1 and F5H2 is induced by wounding. Expression of F5H1 appears to begin earlier than expression of F5H2, and F5H1 expression appears to be reduced in plants with mutation 1, and possibly in plants with mutations in F5H2 gene homologs, such as mutations 3 or 6.

[0085] The present invention relates to plants comprising a modified F5H gene homolog, wherein the gene homolog comprises a modification compared to its corresponding wild-type F5H gene homolog, and wherein the presence of the modified F5H gene homolog in the plant results in reduced wound-induced surface discoloration compared to a plant that does not contain the modified F5H gene homolog.The present invention relates to plants comprising a modified F5H gene of the present invention, wherein the modification results in a premature stop codon.

[0086] The present invention relates to lettuce plants comprising a first modified F5H gene homolog, designated F5H1, the wild type of which has the sequence of SEQ ID NO: 115, and optionally a second modified F5H gene homolog, designated F5H2, the wild type of which has the sequence of SEQ ID NO: 116.

[0087] The present invention also relates to lettuce plants in which a modified F5H1 gene homolog is present in a homozygous form and a modified F5H2 gene homolog is present in either a heterozygous or homozygous form.

[0088] The present invention further relates to a lettuce plant, wherein the modified F5H1 gene comprises a premature stop codon.

[0089] The present invention further relates to lettuce plants, wherein the modified F5H1 gene comprises a premature stop codon caused by a C→T mutation at position 370 of SEQ ID NO:115.

[0090] In one aspect, the invention relates to another plant of the invention listed in Figure 4, wherein the premature stop codon is caused by a mutation at a position corresponding to position 370 of SEQ ID NO: 115 in lettuce.

[0091] The present invention further relates to lettuce plants comprising a first modified F5H gene homolog designated F5H1, the wild-type of which has the sequence of SEQ ID NO: 115, and optionally comprising a second modified F5H gene homolog designated F5H2, the wild-type of which has the sequence of SEQ ID NO: 116, wherein the modified F5H2 gene encodes a protein with one or more amino acid substitutions.

[0092] The present invention further relates to lettuce plants comprising a first modified F5H gene homolog designated F5H1 (the wild-type form of which has the sequence of SEQ ID NO: 115), and optionally comprising a second modified F5H gene homolog designated F5H2 (the wild-type form of which has the sequence of SEQ ID NO: 116), wherein the modified F5H2 gene encodes a protein with an amino acid substitution of threonine to isoleucine at position 154 of the encoded F5H2 protein of SEQ ID NO: 2, and / or an amino acid substitution of glycine to glutamic acid at position 165 of the encoded F5H2 protein of SEQ ID NO: 2, and / or an amino acid substitution of serine to phenylalanine at position 308 of the encoded F5H2 protein of SEQ ID NO: 2, and / or an amino acid substitution of glycine to glutamic acid at position 434 of the encoded F5H2 protein of SEQ ID NO: 2, and / or an amino acid substitution of glycine to glutamic acid at position 436 of the encoded F5H2 protein of SEQ ID NO: 2.

[0093] The present invention further relates to lettuce plants comprising a first modified F5H gene homolog designated F5H1, the wild-type of which has the sequence of SEQ ID NO: 115, and optionally comprising a second modified F5H gene homolog designated F5H2, the wild-type of which has the sequence of SEQ ID NO: 116, wherein an amino acid substitution of threonine to isoleucine at position 154 of the encoded F5H2 protein is the result of a C→T nucleotide change at position 461 of SEQ ID NO: 116, and an amino acid substitution of glycine to glutamic acid at position 165 of the encoded F5H2 protein is the result of a C→T nucleotide change at position 49 of SEQ ID NO: 116. The amino acid substitution of serine to phenylalanine at position 308 of the encoded F5H2 protein is the result of a C to T nucleotide change at position 923 of SEQ ID NO:116, the amino acid substitution of glycine to glutamic acid at position 434 of the encoded F5H2 protein is the result of a G to A nucleotide change at position 1301 of SEQ ID NO:116, and the amino acid substitution of glycine to glutamic acid at position 436 of the encoded F5H2 protein is the result of a G to A nucleotide change at position 1307 of SEQ ID NO:116.

[0094] In one aspect, the invention relates to another plant of the invention listed in Figure 4, wherein the amino acid substitution is at a position corresponding to the position in lettuce.

[0095] The present invention also relates to lettuce plants comprising a modified F5H1 gene containing a premature stop codon and an F5H2 gene containing an amino acid substitution.

[0096] The present invention relates to lettuce plants comprising a modified F5H1 gene comprising a premature stop codon and an F5H2 gene comprising an amino acid substitution, wherein the premature stop codon in the F5H1 gene is caused by a C→T mutation at position 370 of SEQ ID NO:115, the amino acid substitution of threonine to isoleucine in the F5H2 gene at position 154 of the encoded F5H2 protein is the result of a C→T nucleotide change at position 461 of SEQ ID NO:116, and the amino acid substitution of glycine to glutamic acid at position 165 of the encoded F5H2 protein is the result of a G→T nucleotide change at position 494 of SEQ ID NO:116. a serine to phenylalanine amino acid substitution at position 308 of the encoded F5H2 protein is the result of a C→T nucleotide change at position 923 of SEQ ID NO:116; a glycine to glutamic acid amino acid substitution at position 434 of the encoded F5H2 protein is the result of a G→A nucleotide change at position 1301 of SEQ ID NO:116; or a glycine to glutamic acid amino acid substitution at position 436 of the encoded F5H2 protein is the result of a G→A nucleotide change at position 1307 of SEQ ID NO:116.

[0097] In one aspect, the present invention relates to a lettuce plant of the invention comprising a modified F5H1 gene comprising a premature stop codon and an F5H2 gene comprising an amino acid substitution, wherein the premature stop codon in the F5H1 gene is caused by a C→T mutation at position 370 of SEQ ID NO: 115, and the amino acid substitution in the F5H2 gene from glycine to serine at position 159 of the encoded F5H2 protein is the result of a G→A nucleotide change at position 475 of SEQ ID NO: 116.

[0098] In particular, the present invention relates to lettuce plants containing a modification to the F5H1 gene homolog, where the modification, in combination with a modification to the F5H2 gene resulting in an amino acid substitution, leads to a premature stop in the coding sequence of the F5H1 gene of lettuce. When both the mutation leading to a premature stop codon in the F5H1 gene homolog and the amino acid substitution in the F5H2 protein sequence are carried by a plant, the effect is enhanced, and plants containing these mutations exhibit delayed wound-induced surface discoloration. Thus, plants containing modified F5H1 gene homologs and modified F5H2 gene homologs exhibit delayed wound-induced surface discoloration compared to plants containing only the modified F5H1 gene homolog.

[0099] Segregation analysis is performed using F2 plants obtained by crossing a plant that exhibits reduced wound-induced surface discoloration and contains a mutation in the F5H1 gene homolog and a mutation in the F5H2 gene homolog in a heterozygous state with a wild-type plant that does not contain a modified F5H gene homolog. The resulting F1 plants are self-crossed, and the phenotype of F2 plants grown from the resulting seeds is analyzed. The F2 plants contain the mutation(s) in a homozygous state, a heterozygous state, or no mutations. The results of segregation analysis of the traits of the present invention indicate that in order for plants to exhibit reduced wound-induced surface discoloration, a mutation in the F5H1 homolog (C in the F5H1 gene homolog) is required. 370 →T 370 ), preferably in a homozygous state. Plants containing mutation 1 in the F5H1 homolog in a homozygous state and a mutation in the F5H2 gene homolog, for example mutation 3, in a homozygous or heterozygous state, exhibit reduced wound-induced surface discoloration compared to plants containing only mutation 1 in the F5H1 gene homolog.

[0100] In one aspect, the present invention relates to plants comprising mutations to the F5H1 gene homologues of lettuce plants, as shown in Table 3.

[0101] In a further aspect, the present invention relates to a combination of mutation 1 in the F5H1 gene homologue with one or more of mutations 2, 3, 4, 5 and 6 in the F5H2 gene homologue in lettuce plants, as shown in Table 3.

[0102] The present invention further relates to modified F5H gene homologs that confer reduced wound-induced surface discoloration on plants.

[0103] The present invention further relates to the use of modified F5H gene homologs for the development of plants that exhibit reduced wound-induced surface discolouration.

[0104] The modified F5H gene homologs identified in the course of this invention and described herein are not the only modifications to F5H gene homologs that can result in the traits of the invention, and therefore the invention should not be limited to the specific modifications described herein, but extends to all other modifications to genes and / or proteins that result in reduced wound-induced surface discoloration. Using the methods described herein or known in the art, one skilled in the art can introduce the described or other mutations that have the same or similar effect in lettuce or any other plant containing an F5H gene homolog.

[0105] By using the phenotypic screening test described herein, it can be determined whether wound-induced surface discoloration is reduced compared to that of WT plants.Phenotypic testing can be used to detect reduced wound-induced surface discoloration in lettuce and other crops that have F5H gene homologs.The modification of gene homologs that results in reduced wound-induced surface discoloration can be used in any plant that may be affected by discoloration, and is particularly useful in vegetables or fruits.

[0106] Furthermore, those skilled in the art can also detect other F5H gene homologs other than those characterized herein. Those skilled in the art can detect other gene homologs in crops related to the present invention or other crops described herein. After detecting these other homologs, those skilled in the art can modify their sequences using methods described herein or known in the art. Modifying other F5H gene homologs can enhance the reduction of wound-induced surface discoloration.

[0107] Amino acid substitutions can occur in regions of a protein that do not significantly affect the structure, function, and stability of the protein. However, amino acid substitutions occurring at certain positions within well-conserved domains can affect the expression or activity levels of the protein. Multiple sequence alignments between F5H protein orthologs reveal highly conserved positions that may be related to the stability, function, and / or structure of the F5H protein. In accordance with the present invention, F5H protein homologs were identified in all species and found to contain five conserved motifs listed in Table 1 and highlighted in the protein alignment (Figure 4). Non-conservative amino acid changes within these conserved regions can disrupt the stability, function, and / or structure of the encoded F5H protein. However, modifications outside these motifs can also affect the stability, function, and / or structure of the encoded F5H protein.

[0108] More specifically, the G of the F5H2 DNA sequence of lettuce (SEQ ID NO: 116) 1300 G 1301 A 1302 G of the highly conserved glycine residue at position 434 in the lettuce F5H2 protein (SEQ ID NO: 2), encoded by 1300 A 1301 A 1302 In combination with Mutation 1, substitution with a glutamic acid residue encoded by, results in reduced wound-induced surface discoloration to the plant compared to wild-type plants, and results in reduced wound-induced surface discoloration compared to plants containing Mutation 1 alone.

[0109] More specifically, the T of the lettuce F5H2 DNA sequence (SEQ ID NO: 116) 922 C 923 T 924 a highly conserved serine residue at position 308 of the lettuce F5H2 protein (SEQ ID NO: 2), encoded by 922 T 923 T 924 In combination with mutation 1, substitution with a phenylalanine residue encoded by, results in reduced wound-induced surface discoloration to the plant compared to wild-type plants, and results in reduced wound-induced surface discoloration compared to plants containing mutation 1 alone.

[0110] More specifically, the G of the lettuce F5H2 DNA sequence (SEQ ID NO: 116) 1306 G 1307 A 1308 G of the highly conserved glycine residue at position 436 of the lettuce F5H2 protein (SEQ ID NO: 2), encoded by 1306 A 1307 A 1308 In combination with Mutation 1, substitution with a glutamic acid residue encoded by, results in reduced wound-induced surface discoloration to the plant compared to wild-type plants, and results in reduced wound-induced surface discoloration compared to plants containing Mutation 1 alone.

[0111] The present invention is broadly applicable to all plant species and crops that carry at least one functional F5H gene homolog in their genomes. F5H genes present in other plant species are called "gene orthologs" and encode F5H proteins with the same or similar functions. Identification of F5H orthologs, i.e., F5H genes in other species, can be performed by many methods known in the art. In this study, orthologs of the F5H gene were identified in other crops by comparing the lettuce F5H protein sequence (SEQ ID NOS: 1 and 2) with sequences from other plant genomes using the Basic Local Alignment Search Tool (BLAST) program. The best hits per species were identified as candidate F5H orthologous genes, listed in Table 2. Multiple sequence alignment of the protein sequences using CLUSTAL confirmed that the candidate genes were orthologous F5H genes (Figure 4). Once the DNA sequences of the orthologous F5H genes and their encoded F5H proteins are known, this information can be used to regulate or modify the proteins encoded by the genes using methods described herein or known to those of skill in the art.

[0112] Accordingly, the present invention also relates to the plant species potato, onion, artichoke, rice, corn, peach, eggplant, chicory, endive, celery, apple, banana, soybean, pear, wheat, radish and cabbage, which contain one or more modified F5H gene homologs in their genome, wherein the modified homologs result in reduced wound-induced surface discolouration.

[0113] The present invention relates to a method for selecting plants that exhibit reduced wound-induced surface discolouration, the method comprising screening a plant or a population of plants for the presence of a modified F5H gene homolog that results in reduced wound-induced surface discolouration in the plant, optionally applying a phenotypic test to identify plants that exhibit reduced wound-induced surface discolouration, and selecting plants that exhibit reduced wound-induced surface discolouration.

[0114] Methods used to detect and select plants that exhibit reduced wound-induced discolouration are, for example, phenotypic tests such as those described and illustrated in Example 2, and / or the use of molecular markers as characterized in Example 3. Both methods can be used to directly or indirectly detect and select plants that exhibit reduced wound-induced surface discolouration and contain a modified F5H gene in the F1 or any further generations resulting from crosses with a parent plant that exhibits reduced wound-induced discolouration and contains a modified F5H gene homolog.

[0115] The trait of the present invention may be, for example, a phenotype determined in the leaf disc test described in Example 2, which involves inducing damage by harvesting a plant leaf disc. The shape of the sample is not limited to a disc, but rather any portion of a leaf bearing damage, regardless of shape. The leaf sample is incubated between wet filter paper moistened with MES buffer, and after 3, 5, and 10 days of incubation at 7.5°C, it is compared with leaf samples harvested from control plants, i.e., wild-type plants, that do not carry a modified F5H gene homolog in their genomes and are incubated under the same conditions for the same period.

[0116] The presence and intensity of wound-induced surface discoloration on different leaf disc samples can be evaluated using an appropriate scale for comparison. Those skilled in the art can use any refined scale. In the phenotypic analysis described in Example 2, wound-induced surface discoloration appears as a pink ring around the edge of the leaf disc. When the color is saturated and the wound-induced surface discoloration is significantly strong, the discoloration can range from red to very dark red. An example scale is 9 to 0, where 9 means no discoloration on the edge, a score of 8 means the leaf disc has very slight pink discoloration around the edge, a score of 5 means the leaf disc has a light red / pink discoloration ring around the edge, a score of 2 means the leaf disc has a darker and more intense red / pink discoloration ring around the edge compared to leaf discs with scores of 9, 8, 7, 6, 5, 4, or 3, and a score of 0 means the leaf disc has a significantly darker red, intense discoloration ring around the edge. Examples of each score are shown in Figure 5. The scale described herein is an example of a scale that can be used to assign a score to the leaf discs of the tested plant to compare them with other plants and to identify reduced damage-induced surface discoloration. Preferably, the leaf discs are scored by one person.

[0117] To identify reduced wound-induced surface discoloration, the scores of the tested plants should be compared with those of wild-type plants, for example, 3, 5, and 10 days after sampling. Leaf discs collected from plants exhibiting reduced wound-induced surface discoloration have a higher score than leaf discs from wild-type plants on the same day of incubation. Preferably, leaf discs collected from plants exhibiting reduced wound-induced surface discoloration have a score of 9 or 8. Plants containing modified F5H1 and modified F5H2 genes have a higher score than plants containing only modified F5H1 gene homologs on the same day of incubation. To enable comparison, plant leaf disc tests should be performed under the same conditions using plants grown under the same conditions.

[0118] Alternatively, wound-induced surface discoloration can also be determined by cutting a plant or a portion thereof and storing it until the cut portion exhibits wound-induced surface discoloration. Wound-induced surface discoloration can be evaluated by replicating the plant's normal storage conditions. Mature plants are harvested and cut into pieces. For lettuce plants, the cutting method may vary depending on the lettuce variety used to conduct the test. The plant pieces are washed and stored in a plastic bag in a cool cell at 5-6°C. At 1, 2, 3, 4, 7, and 10 days after washing, the presence and intensity of wound-induced surface discoloration are evaluated. To detect discoloration, the cut leaf pieces were compared with cut leaf pieces from WT plants. The presence and intensity of wound-induced surface discoloration can be detected by the presence and intensity of pink discoloration at the cut ends, which can be evaluated using a 9 to 4 scale, where 9 means no signs of wound-induced surface discoloration, 8 means the first traces of pink discoloration (barely visible, slight redness), 7 means pink discoloration is present at some ends, 6 means pink discoloration is present at all cut ends, 5 means strong pink discoloration is present at all cut ends, and 4 means deep pink discoloration is present at all cut ends. Plants showing reduced wound-induced surface discoloration have a higher score than wild-type plants. To ensure comparability, phenotypic analysis of whole lettuce heads should be performed under the same conditions using plants grown under the same conditions.

[0119] The present invention relates to a molecular marker for detecting a mutation in the genome of a plant that causes reduced wound-induced surface discolouration in said plant or part thereof, said molecular marker being a mutation in any of the wild-type sequences of the SEQ ID NOs shown in Table 2.

[0120] The present invention relates to a molecular marker for detecting a mutation in the genome of a plant that causes reduced wound-induced surface discoloration in the plant or a part thereof, wherein the mutation is a C→T nucleotide change at position 370 of SEQ ID NO: 115.

[0121] The present invention relates to a molecular marker for detecting a mutation in the genome of a plant that causes reduced wound-induced surface discolouration in the plant or a part thereof, wherein the mutation is a C→T mutation at position 461 of SEQ ID NO: 116, resulting in an amino acid substitution from threonine to isoleucine at position 154 of the encoded protein, and / or a G→A mutation at position 494 of SEQ ID NO: 116, resulting in an amino acid substitution from glycine to glutamic acid at position 165 of the encoded protein, and / or a C→T mutation at position 923 of SEQ ID NO: 116, resulting in an amino acid substitution from serine to phenylalanine at position 308 of the encoded protein, and / or a G→A mutation at position 1301 of SEQ ID NO: 116, resulting in an amino acid substitution from glycine to glutamic acid at position 434 of the encoded protein, and / or a G→A mutation at position 1307 of SEQ ID NO: 116, resulting in an amino acid substitution from glycine to glutamic acid at position 436 of the encoded protein.

[0122] The present invention further relates to the use of the molecular markers described herein to identify or develop plants that exhibit reduced wound-induced surface discolouration, or to develop other markers linked to the modified F5H gene homologs of the present invention.

[0123] The present invention also provides a method for identifying molecular markers associated with reduced wound-induced surface discolouration in plants, comprising: a) isolating DNA from a plant and one or both parents of the plant; b) screening for molecular markers in the region of said DNA at or near the sequence corresponding to SEQ ID NO: 174, 175, 176, 177, 178 or 179; and c) determining the co-inheritance of said markers with the reduced wound-induced discolouration phenotype from one or both parents of said plant. The present invention also relates to a method, including:

[0124] The present invention also relates to the use of molecular markers to identify modifications in F5H gene homologs that result in reduced wound-induced surface discoloration, or to develop other markers related to the modified F5H gene homologs of the present invention. Molecular markers are based on modifications to the F5H gene homologs that underlie the trait. A non-exclusive list of suitable molecular markers is provided herein. Those skilled in the art are familiar with creating and using them to detect and select plants with modified F5H gene homologs that cause reduced wound-induced surface discoloration during breeding.

[0125] During the research leading to the present invention, numerous EMS-induced SNP mutations were identified in two F5H gene homologs in lettuce. One of the SNPs identified in F5H1 resulted in a stop codon in the protein, and five SNPs identified in F5H2 resulted in amino acid changes in the protein sequence. SNPs can be used as markers to detect the presence of modified F5H gene homologs in plant genomes.

[0126] In a particular embodiment, one suitable molecular marker is the C5H1 gene in the F5H1 gene of lettuce (Lactuca sativa) shown in Table 5 (Example 3). 370 →T 370 It is a SNP.

[0127] In a particular embodiment, one suitable molecular marker is the C in the F5H2 gene of lettuce (Lactuca sativa) shown in Table 6 (Example 3). 461 →T 461 It is a SNP.

[0128] In a particular embodiment, one suitable molecular marker is the G in the F5H2 gene of lettuce (Lactuca sativa) shown in Table 6 (Example 3).494 →A 494 It is a SNP.

[0129] In a particular embodiment, one suitable molecular marker is the C in the F5H2 gene of lettuce (Lactuca sativa) shown in Table 6 (Example 3). 923 →T 923 It is a SNP.

[0130] In a particular embodiment, one suitable molecular marker is the G in the F5H2 gene of lettuce (Lactuca sativa) shown in Table 6 (Example 3). 1301 →A 1301 It is a SNP.

[0131] In a particular embodiment, one suitable molecular marker is the G in the F5H2 gene of lettuce (Lactuca sativa) shown in Table 6 (Example 3). 1307 →A 1307 It is a SNP.

[0132] The present invention relates to molecular markers and the use of these markers to identify modified F5H gene homologs that result in reduced wound-induced surface discolouration in all plants containing an F5H gene homolog. The above SNP markers are particularly suitable for use in lettuce.

[0133] The present invention relates to a method for determining the presence of a modified F5H gene homolog in a plant of the invention, comprising the steps of obtaining a nucleic acid sample from the plant, comparing the nucleic acid to a nucleic acid sample obtained from a control plant comprising a wild-type F5H gene homolog, and detecting a polymorphism between the two nucleic acid samples, wherein the detected polymorphism is indicative of the presence of the modified homolog.

[0134] Preferably, the wild-type F5H gene homolog is any one of the sequences listed in Table 2.

[0135] The modified F5H gene can be introduced into other genetic backgrounds, either of the same species or different species. Plants lacking the modification may have other desired traits. For sexually compatible plants, gene transfer can be achieved by crossing and / or backcrossing and selecting in the first generation where reduced wound-induced surface discoloration is detectable. Crossing can be supplemented, if necessary, with embryo rescue techniques or other techniques that result in successful combination and gene transfer; these techniques are known to those skilled in the art. The parent plant can be a plant directly cultivated from the deposited seed or a progeny plant derived from the seed or a progeny plant derived from a seed identified by other means as having the trait of the present invention.

[0136] When a trait is dominant and single-gene, it can be transferred into another plant in just one generation (F1). When the trait is recessive and / or involves more than one gene, transfer can involve a breeding process requiring multiple generations. Transfer is used herein to describe the entire process. For dominant traits, selection of plants carrying the modification can begin with F1, or with further generations obtained by crossing a plant with the desired trait with a plant that does not have the trait. For recessive traits, selection by phenotypic testing and / or selection using molecular markers can begin with F2, or with further generations obtained from crossing or another generation obtained from backcrossing.

[0137] In certain embodiments, one or more modified lettuce F5H gene homologs can be genetically transferred from a lettuce plant carrying a modified lettuce F5H gene homolog into a lettuce plant lacking the modified lettuce F5H gene homolog using standard breeding techniques.

[0138] The present invention further relates to a seedling suitable for producing plants containing one or more modified F5H genes in its genome and exhibiting reduced wound-induced discoloration. In one embodiment, the seedling is formed from plant parts suitable for sexual reproduction, particularly microspores, pollen, ovaries, ovules, embryos, embryo sacs, and egg cells. In another embodiment, the seedling is formed from plant parts suitable for vegetative propagation, particularly microspores, pollen, ovaries, ovules, embryos, embryo sacs, egg cells, cuttings, roots, root tips, hypocotyls, cotyledons, stems, leaves, flowers, anthers, seeds, meristematic cells, protoplasts, and cells, or tissue cultures thereof.

[0139] The present invention also relates to plants grown or regenerated from said seedlings, which plants comprise in their genome one or more modified F5H genes as defined herein, which provide said plants with reduced wound-induced surface discolouration.

[0140] In particular, plants produced from the seedlings contain a modified F5H gene homologue as found in lettuce plants grown from seed, and representative seeds have been deposited with NCIMB under accession numbers NCIMB42546, NCIMB42547, NCIMB42548, NCIMB42549, NCIMB42550 and NCIMB42551.

[0141] The present invention also relates to the use of plants of the present invention that contain a modified F5H gene that provides the plant with reduced wound-induced surface discolouration in plant breeding to confer this trait.

[0142] The present invention provides a method for producing a plant that exhibits reduced wound-induced surface discolouration, comprising the steps of: a) crossing a plant containing the modified F5H gene homolog of claim 1 with another plant; b) optionally, performing one or more selfing and / or outcrossings; and c) optionally selecting after each round of self-crossing or out-crossing for plants containing reduced wound-induced surface discoloration. The present invention relates to a method comprising:

[0143] In one embodiment, plants are selected phenotypically and / or by the use of molecular markers.

[0144] In one aspect, the present invention provides a method for producing a plant that exhibits reduced wound-induced surface discoloration, comprising the steps of: a) crossing a plant containing a modified F5H gene homolog of the present invention that confers the trait with another plant; b) self-crossing the resulting F1 to obtain F2 plants; c) selecting plants in the F2 that have the trait; d) optionally, one or more rounds of self- or out-crossing and subsequent selection for plants containing / exhibiting the trait of the invention. The present invention relates to a method comprising:

[0145] In one aspect, the present invention provides a method for producing a plant that exhibits reduced wound-induced surface discoloration, comprising the steps of: a) crossing a plant containing a modified F5H gene homolog of the present invention that confers the trait with another plant; b) optionally backcrossing the resulting F1 with a preferred parent; c) selecting plants having the trait in the F2; d) optionally, one or more rounds of self- or out-crossing and subsequent selection for plants containing the trait; The present invention relates to a method comprising:

[0146] The present invention further provides a method for introducing another desired trait into a plant having a trait of the present invention, comprising the steps of: a) crossing a plant containing a modified F5H gene homolog of the present invention and exhibiting reduced wound-induced surface discoloration with a plant containing the desired trait to produce F1 progeny; b) selecting F1 progeny plants that contain the reduced wound-induced surface discoloration trait and a desired trait; c) mating the selected F1 progeny with either parent to produce backcrossed progeny; d) selecting backcrossed progeny that contain the desired traits and exhibit reduced wound-induced surface discoloration; and e) optionally repeating steps c) and d) one or more successive times to generate selected fourth or more backcrossed progeny that contain the modified F5H gene homolog and that exhibit reduced wound-induced surface discoloration. The present invention includes plants produced by this method.

[0147] In one embodiment, selection for plants exhibiting reduced wound-induced surface discoloration is performed in the F1 or in additional generations using the markers described in Example 3. In another aspect, selection for the traits of the invention begins in the F2 of the cross or in another generation of the backcross. Selection of plants in the F2 can be performed phenotypically as well as by using the marker(s) that directly or indirectly detect the modified F5H gene underlying the trait.

[0148] In one embodiment, selection for plants exhibiting reduced wound-induced surface discolouration begins in the F3 or later generation.

[0149] In one embodiment, a plant comprising an F5H gene homolog of the present invention is a self-cross line, a hybrid, a doubled haploid, or a segregating population plant.

[0150] The invention further provides methods for producing plants that exhibit reduced wound-induced surface discolouration using doubled haploid production techniques to produce doubled haploid lines containing the trait.

[0151] The present invention further relates to hybrid seeds that can be grown into plants that exhibit reduced wound-induced surface discoloration and methods for producing such hybrid seeds, comprising crossing a first parent plant with a second parent plant and harvesting the resulting seeds, wherein the first parent plant and / or the second parent plant is a plant of the present invention.

[0152] The present invention further relates to a method for producing a hybrid plant that exhibits reduced wound-induced surface discoloration, the method comprising crossing a first parent plant with a second parent plant and harvesting the resulting hybrid seed, wherein the first parent plant and / or the second parent plant is a plant that exhibits reduced wound-induced surface discoloration, and growing the hybrid seed into a hybrid plant that exhibits reduced wound-induced surface discoloration.

[0153] The present invention also relates to a method for producing a hybrid plant seed, comprising crossing a first parent plant with a second parent plant and harvesting the resulting plant seed, wherein the first parent plant and / or the second parent plant comprises a modified F5H gene homolog of the present invention.

[0154] The present invention also relates to a method of producing plants that exhibit reduced wound-induced surface discolouration, comprising growing plants derived from seeds that contain within their genome a modified F5H gene homolog that confers the trait of reduced wound-induced surface discolouration, preferably the seeds of which representative samples have been deposited with NCIMB under accession numbers NCIMB42546, NCIMB42547, NCIMB42548, NCIMB42549, NCIMB42550 and NCIMB42551.

[0155] The present invention also relates to a method of seed production comprising growing plants from seeds, representative samples of which have been deposited with NCIMB under accession numbers NCIMB42546, NCIMB42547, NCIMB42548, NCIMB42549, NCIMB42550 and NCIMB42551, thereby allowing the plants to produce seeds, and harvesting the seeds. Seed production is preferably by outcrossing or self-crossing.

[0156] The present invention relates to methods for producing plants that exhibit reduced wound-induced surface discolouration by tissue culture using the plants of the invention described herein as a source of tissue.

[0157] The present invention further relates to methods for producing plants that exhibit reduced wound-induced surface discolouration by vegetative propagation of parts of the plants of the invention described herein.

[0158] In one aspect, the present invention relates to a method for producing a plant that exhibits reduced wound-induced surface discoloration by using a method for genetic modification to introduce the trait from a plant of the present invention into a plant that exhibits said reduced wound-induced surface discoloration. Genetic modification includes transgenic modification or genetic recombination, using genes from non-cross-breeding species or synthetic genes, as well as cisgenic modification or cisgenesis, using native genes encoding (agronomic) traits from the plant itself or from a sexually compatible donor plant.

[0159] The present invention also relates to breeding methods for developing plants that exhibit reduced wound-induced surface discolouration, wherein germplasm containing the trait is used. Representative seeds and representative germplasm of the plants containing modified F5H gene homologs have been deposited with NCIMB under accession numbers NCIMB42546, NCIMB42547, NCIMB42548, NCIMB42549, NCIMB42550 and NCIMB42551.

[0160] In a further aspect, the present invention relates to a plant that exhibits reduced wound-induced surface discolouration, wherein progeny or seedlings of a plant containing a modified F5H gene homolog that confers the trait are used as a source for introducing the trait into another plant. Representative seeds of the plants containing modified F5H gene homologs have been deposited with NCIMB under accession numbers NCIMB42546, NCIMB42547, NCIMB42548, NCIMB42549, NCIMB42550 and NCIMB42551.

[0161] The present invention preferably provides plants that exhibit reduced wound-induced surface discolouration, which plants can be obtained using any of the methods described herein and / or known to those skilled in the art.

[0162] In the process of breeding new plants carrying a modified F5H gene homolog, desired agronomic traits can be introduced into the plant independently of the modified F5H gene. As used herein, "desired traits" include, but are not limited to, improved yield, leaf shape, leaf size, leaf number, leaf color, seed number, seed size, plant vigor, plant height, bolting, and resistance to one or more diseases or diseases caused by organisms. Any one of these desired traits can be combined with a modified F5H gene homolog.

[0163] The present invention further relates to a method for producing an agronomically elite plant of the present invention that exhibits reduced wound-induced surface discoloration, comprising introducing a modified F5H gene homolog into an agronomically elite plant, which may be achieved by methods described herein or known to those skilled in the art. The present invention also includes plants produced by this method.

[0164] In still further embodiments, the agronomically elite plants of the present invention are inbred lines or hybrids.

[0165] As used herein, an inbred plant is a plant from a plant population resulting from three or more self-crosses or backcrosses; or is a doubled haploid plant. Inbred plants can be, for example, parental lines used in the production of commercial hybrid varieties.

[0166] As used herein, hybrid plant refers to the plant obtained by crossing two different plants with different genotypes.More specifically, hybrid plant is obtained by crossing two different self-crossing lineage plants, and such hybrid plant can be, for example, F1 hybrid variety plant.

[0167] In one embodiment, the plants of the present invention, ie, plants comprising a modified F5H gene of the present invention, are agriculturally elite plants.

[0168] As used herein, an agriculturally elite plant is a plant that has a genotype that, as a result of breeding and selection directed by human intervention, results in an accumulation of distinguishable and desirable agronomic traits that enable growers to harvest commercially meaningful products.

[0169] The present invention also relates to food products comprising parts of the plants of the present invention. The food products may include one or more harvested parts of the plants of the present invention, and the present invention relates to food products that may include harvested leaves of the plants of the present invention, either in natural or processed form, and containers that may contain one or more plants of the present invention in a growing substrate for harvesting leaves from lettuce plants in a domestic environment. The harvested parts or food products may be or include the heads and / or parts of the plants of the present invention, such as leaves. The food products or harvested parts may have undergone one or more processing steps. Such processing steps may include, but are not limited to, any one or a combination of the following treatments: cutting, washing, or salad mixes that may include parts of the plants of the present invention, such as leaves. The resulting processed forms are also part of the present invention. All food products and harvested parts retain their genomically modified F5H gene homologs of the present invention.

[0170] The present invention relates to a part of a plant of the invention, wherein said part is a leaf, the whole head of the plant, a fruit, an inflorescence, a seed, a curd, a stem, a tuber, a bulb or a root, optionally in treated form.

[0171] The present invention further relates to seeds that are capable of growing into plants of the present invention.

[0172] The present invention also relates to seeds of the plants of the present invention, wherein the seeds comprise a modified F5H gene homologue in their genome.

[0173] The present invention further relates to a cell of a plant of the present invention, which comprises within its genome a modified F5H gene and provides the plant with reduced wound-induced discolouration. Such a cell may be isolated from or be part of a plant or part thereof.

[0174] The present invention also relates to cells of lettuce plants that exhibit reduced wound-induced surface discolouration, as seen in lettuce plants (Lactuca sativa) grown from seeds deposited with NCIMB on February 19, 2016, under one of the accession numbers NCIMB42546, NCIMB42547, NCIMB42548, NCIMB42549, NCIMB42550 or NCIMB42551.

[0175] The present invention also relates to plant cells that contain a modified F5H gene and that exhibit reduced wound-induced surface discolouration. The present invention also relates to plant cells that contain a modified F5H gene and that exhibit reduced wound-induced surface discoloration, wherein the plants are obtained by crossing plants that contain a modified F5H gene and selecting for plants that exhibit reduced wound-induced surface discoloration.

[0176] The present invention also relates to cells of a lettuce plant that exhibits reduced wound-induced surface discolouration, as found in lettuce plants (Lactuca sativa) grown from seeds deposited with NCIMB on February 19, 2016 under one of accession numbers NCIMB42546, NCIMB42547, NCIMB42548, NCIMB42549, NCIMB42550 or NCIMB42551, wherein the lettuce plant may be obtained by crossing a lettuce plant with a lettuce plant grown from seeds deposited with NCIMB on February 19, 2016 under one of accession numbers NCIMB42546, NCIMB42547, NCIMB42548, NCIMB42549, NCIMB42550 or NCIMB42551, and selecting for a lettuce plant that exhibits reduced wound-induced surface discolouration.

[0177] The present invention also relates to the use of seeds deposited with NCIMB under accession numbers NCIMB42546, NCIMB42547, NCIMB42548, NCIMB42549, NCIMB42550 and NCIMB42551 for growing plants that exhibit reduced wound-induced surface discolouration in another lettuce plant (Lactuca sativa).

[0178] The present invention also relates to the use of seeds of representative samples deposited with NCIMB under accession numbers NCIMB42546, NCIMB42547, NCIMB42548, NCIMB42549, NCIMB42550 and NCIMB42551 to transfer the reduced wound-induced surface discolouration trait to another agronomically valuable lettuce plant.

[0179] The present invention also relates to the use of plants comprising a modified F5H gene homologue of the present invention and exhibiting reduced wound-induced surface discolouration as crop plants.

[0180] In particular, the present invention relates to the use of lettuce plants (Lactuca sativa) as a crop plant that exhibits reduced wound-induced surface discolouration, as seen in lettuce plants grown from seeds deposited with NCIMB on February 19, 2016 under one of the accession numbers NCIMB42546, NCIMB42547, NCIMB42548, NCIMB42549, NCIMB42550 or NCIMB42551.

[0181] The present invention also relates to the use of plants comprising a modified F5H gene homologue of the present invention and exhibiting reduced wound-induced surface discolouration as a source of seeds.

[0182] In particular, the present invention relates to the use of lettuce plants (Lactuca sativa) as a seed source that exhibit reduced wound-induced surface discolouration, as seen in lettuce plants grown from seeds deposited with NCIMB on February 19, 2016 under one of the accession numbers NCIMB42546, NCIMB42547, NCIMB42548, NCIMB42549, NCIMB42550 or NCIMB42551.

[0183] The present invention also relates to the use of plants comprising a modified F5H gene homolog of the present invention and exhibiting reduced wound-induced surface discolouration as a seed source.

[0184] In particular, the present invention relates to the use of lettuce plants (Lactuca sativa) that exhibit reduced wound-induced surface discolouration, as seen in lettuce plants grown from seeds deposited with NCIMB on February 19, 2016 under one of the accession numbers NCIMB42546, NCIMB42547, NCIMB42548, NCIMB42549, NCIMB42550 or NCIMB42551, as a seed source.

[0185] The present invention also relates to the use of plants comprising a modified F5H gene homologue of the present invention and exhibiting reduced wound-induced surface discolouration for food consumption.

[0186] In particular, the present invention relates to the use, for consumption, of lettuce plants (Lactuca sativa) that exhibit reduced wound-induced surface discolouration, as seen in lettuce plants grown from seeds deposited with NCIMB on February 19, 2016 under one of the accession numbers NCIMB42546, NCIMB42547, NCIMB42548, NCIMB42549, NCIMB42550 or NCIMB42551. The present invention also relates to the use of a modified F5H gene homolog of the present invention to provide a plant with reduced wound-induced surface discolouration.

[0187] In particular, the present invention relates to the use of an F5H gene homolog, such as that found in seeds deposited with NCIMB on February 19, 2016 under one of the accession numbers NCIMB42546, NCIMB42547, NCIMB42548, NCIMB42549, NCIMB42550 or NCIMB 42551, to provide lettuce plants (Lactuca sativa) with reduced wound-induced surface discolouration.

[0188] The present invention relates to the use of plants as recipients of the modified F5H gene homologues of the present invention.

[0189] In particular, the present invention relates to the use of lettuce plants (Lactuca sativa) as found in the seeds deposited with NCIMB on February 19, 2016 under one of the accession numbers NCIMB42546, NCIMB42547, NCIMB42548, NCIMB42549, NCIMB42550 or NCIMB42551.

[0190] The present invention also relates to the use of a modified F5H gene of the present invention to provide a plant with reduced wound-induced surface discolouration.

[0191] In particular, the present invention relates to the use of a modified F5H gene homolog, as found in seeds deposited with NCIMB on February 19, 2016 under one of the accession numbers NCIMB42546, NCIMB42547, NCIMB42548, NCIMB42549, NCIMB42550 or NCIMB42551, to provide lettuce plants (Lactuca sativa) with reduced wound-induced surface discolouration.

[0192] The present invention also relates to the use of seeds deposited with NCIMB on February 19, 2016 under one of the accession numbers NCIMB42546, NCIMB42547, NCIMB42548, NCIMB42549, NCIMB42550 or NCIMB42551 to transfer reduced wound-induced surface discolouration to another lettuce plant (Lactuca sativa).

[0193] Deposit information Seeds of lettuce plants (Lactuca sativa) of the present invention containing modified F5H gene homologs that confer reduced wound-induced surface discolouration were deposited on February 19, 2016 with NCIMB Ltd. (Ferguson Building, Craibstone Estate, Bucksburn, Aberdeen AB21 9YA, UK) under accession numbers NCIMB42546, NCIMB42547, NCIMB42548, NCIMB42549, NCIMB42550 and NCIMB42551.

[0194] The deposited seeds do not meet the DUS criteria required to obtain plant variety protection and therefore cannot be considered a plant variety. [Brief explanation of the drawings]

[0195] In the examples, reference is made to the following figures:

[0196] [Figure 1] Figure 1: Leaf discs of wild-type (WT) and various mutant lettuce (Lactuca sativa) samples from the phenotypic test described in Example 2 on day 3 of incubation. Lanes 1 and 8 are leaf disc samples taken from wild-type lettuce plants, lanes 2 and 9 are leaf disc samples taken from lettuce plants containing mutation 1, lane 3 is a leaf disc sample taken from lettuce plants containing mutations 1 and 2, lane 4 is a leaf disc sample taken from lettuce plants containing mutations 1 and 3, lane 5 is a leaf disc sample taken from lettuce plants containing mutations 1 and 4, lane 6 is a leaf disc sample taken from lettuce plants containing mutations 1 and 5, and lane 7 is a leaf disc sample taken from lettuce plants containing mutations 1 and 6. The mutants are listed in Table 3. [Figure 2]Figure 2: Leaf discs of wild-type (WT) and various mutant lettuce (Lactuca sativa) samples from the phenotypic test described in Example 2 on day 5 of incubation. Lanes 1 and 8 are leaf disc samples taken from wild-type lettuce plants, lanes 2 and 9 are leaf disc samples taken from lettuce plants containing mutation 1, lane 3 is a leaf disc sample taken from lettuce plants containing mutations 1 and 2, lane 4 is a leaf disc sample taken from lettuce plants containing mutations 1 and 3, lane 5 is a leaf disc sample taken from lettuce plants containing mutations 1 and 4, lane 6 is a leaf disc sample taken from lettuce plants containing mutations 1 and 5, and lane 7 is a leaf disc sample taken from lettuce plants containing mutations 1 and 6. The mutants are listed in Table 3. [Figure 3] Figure 3: Leaf discs of wild-type (WT) and various mutant lettuce (Lactuca sativa) samples from the phenotypic test described in Example 2 on day 10 of incubation. Lanes 1 and 8 are leaf disc samples taken from wild-type lettuce plants, lanes 2 and 9 are leaf disc samples taken from lettuce plants containing mutation 1, lane 3 is a leaf disc sample taken from lettuce plants containing mutations 1 and 2, lane 4 is a leaf disc sample taken from lettuce plants containing mutations 1 and 3, lane 5 is a leaf disc sample taken from lettuce plants containing mutations 1 and 4, lane 6 is a leaf disc sample taken from lettuce plants containing mutations 1 and 5, and lane 7 is a leaf disc sample taken from lettuce plants containing mutations 1 and 6. The mutants are listed in Table 3. [Figure 4-1]Figure 4: Alignment of orthologous proteins with the wild-type sequences of the SEQ ID NOs listed in Table 2 (CLUSTAL multiple sequence alignment with MUSCLE (3.8)). The motifs from Table 1 are highlighted in the alignment. The following symbols are used in the alignment: * -- all residues in that column are identical: -- conservative substitutions observed. -- semi-conservative substitutions observed. -- no match (space). [Figure 4-2] Figure 4: Alignment of orthologous proteins with the wild-type sequences of the SEQ ID NOs listed in Table 2 (CLUSTAL multiple sequence alignment with MUSCLE (3.8)). The motifs from Table 1 are highlighted in the alignment. The following symbols are used in the alignment: * -- all residues in that column are identical: -- conservative substitutions observed. -- semi-conservative substitutions observed. -- no match (space). [Figure 4-3] Figure 4: Alignment of orthologous proteins with the wild-type sequences of the SEQ ID NOs listed in Table 2 (CLUSTAL multiple sequence alignment with MUSCLE (3.8)). The motifs from Table 1 are highlighted in the alignment. The following symbols are used in the alignment: * -- all residues in that column are identical: -- conservative substitutions observed. -- semi-conservative substitutions observed. -- no match (space). [Figure 4-4] Figure 4: Alignment of orthologous proteins with the wild-type sequences of the SEQ ID NOs listed in Table 2 (CLUSTAL multiple sequence alignment with MUSCLE (3.8)). The motifs from Table 1 are highlighted in the alignment. The following symbols are used in the alignment: * -- all residues in that column are identical: -- conservative substitutions observed. -- semi-conservative substitutions observed. -- no match (space). [Figure 4-5]Figure 4: Alignment of orthologous proteins with the wild-type sequences of the SEQ ID NOs listed in Table 2 (CLUSTAL multiple sequence alignment with MUSCLE (3.8)). The motifs from Table 1 are highlighted in the alignment. The following symbols are used in the alignment: * -- all residues in that column are identical: -- conservative substitutions observed. -- semi-conservative substitutions observed. -- no match (space). [Figure 4-6] Figure 4: Alignment of orthologous proteins with the wild-type sequences of the SEQ ID NOs listed in Table 2 (CLUSTAL multiple sequence alignment with MUSCLE (3.8)). The motifs from Table 1 are highlighted in the alignment. The following symbols are used in the alignment: * -- all residues in that column are identical: -- conservative substitutions observed. -- semi-conservative substitutions observed. -- no match (space). [Figure 4-7] Figure 4: Alignment of orthologous proteins with the wild-type sequences of the SEQ ID NOs listed in Table 2 (CLUSTAL multiple sequence alignment with MUSCLE (3.8)). The motifs from Table 1 are highlighted in the alignment. The following symbols are used in the alignment: * -- all residues in that column are identical: -- conservative substitutions observed. -- semi-conservative substitutions observed. -- no match (space). [Figure 4-8] Figure 4: Alignment of orthologous proteins with the wild-type sequences of the SEQ ID NOs listed in Table 2 (CLUSTAL multiple sequence alignment with MUSCLE (3.8)). The motifs from Table 1 are highlighted in the alignment. The following symbols are used in the alignment: * -- all residues in that column are identical: -- conservative substitutions observed. -- semi-conservative substitutions observed. -- no match (space). [Figure 4-9]Figure 4: Alignment of orthologous proteins with the wild-type sequences of the SEQ ID NOs listed in Table 2 (CLUSTAL multiple sequence alignment with MUSCLE (3.8)). The motifs from Table 1 are highlighted in the alignment. The following symbols are used in the alignment: * -- all residues in that column are identical: -- conservative substitutions observed. -- semi-conservative substitutions observed. -- no match (space). [Figure 4-10] Figure 4: Alignment of orthologous proteins with the wild-type sequences of the SEQ ID NOs listed in Table 2 (CLUSTAL multiple sequence alignment with MUSCLE (3.8)). The motifs from Table 1 are highlighted in the alignment. The following symbols are used in the alignment: * -- all residues in that column are identical: -- conservative substitutions observed. -- semi-conservative substitutions observed. -- no match (space). [Figure 4-11] Figure 4: Alignment of orthologous proteins with the wild-type sequences of the SEQ ID NOs listed in Table 2 (CLUSTAL multiple sequence alignment with MUSCLE (3.8)). The motifs from Table 1 are highlighted in the alignment. The following symbols are used in the alignment: * -- all residues in that column are identical: -- conservative substitutions observed. -- semi-conservative substitutions observed. -- no match (space). [Figure 4-12] Figure 4: Alignment of orthologous proteins with the wild-type sequences of the SEQ ID NOs listed in Table 2 (CLUSTAL multiple sequence alignment with MUSCLE (3.8)). The motifs from Table 1 are highlighted in the alignment. The following symbols are used in the alignment: * -- all residues in that column are identical: -- conservative substitutions observed. -- semi-conservative substitutions observed. -- no match (space). [Figure 4-13]Figure 4: Alignment of orthologous proteins with the wild-type sequences of the SEQ ID NOs listed in Table 2 (CLUSTAL multiple sequence alignment with MUSCLE (3.8)). The motifs from Table 1 are highlighted in the alignment. The following symbols are used in the alignment: * -- all residues in that column are identical: -- conservative substitutions observed. -- semi-conservative substitutions observed. -- no match (space). [Figure 4-14] Figure 4: Alignment of orthologous proteins with the wild-type sequences of the SEQ ID NOs listed in Table 2 (CLUSTAL multiple sequence alignment with MUSCLE (3.8)). The motifs from Table 1 are highlighted in the alignment. The following symbols are used in the alignment: * -- all residues in that column are identical: -- conservative substitutions observed. -- semi-conservative substitutions observed. -- no match (space). [Figure 4-15] Figure 4: Alignment of orthologous proteins with the wild-type sequences of the SEQ ID NOs listed in Table 2 (CLUSTAL multiple sequence alignment with MUSCLE (3.8)). The motifs from Table 1 are highlighted in the alignment. The following symbols are used in the alignment: * -- all residues in that column are identical: -- conservative substitutions observed. -- semi-conservative substitutions observed. -- no match (space). [Figure 4-16] Figure 4: Alignment of orthologous proteins with the wild-type sequences of the SEQ ID NOs listed in Table 2 (CLUSTAL multiple sequence alignment with MUSCLE (3.8)). The motifs from Table 1 are highlighted in the alignment. The following symbols are used in the alignment: * -- all residues in that column are identical: -- conservative substitutions observed. -- semi-conservative substitutions observed. -- no match (space). [Figure 4-17]Figure 4: Alignment of orthologous proteins with the wild-type sequences of the SEQ ID NOs listed in Table 2 (CLUSTAL multiple sequence alignment with MUSCLE (3.8)). The motifs from Table 1 are highlighted in the alignment. The following symbols are used in the alignment: * -- all residues in that column are identical: -- conservative substitutions observed. -- semi-conservative substitutions observed. -- no match (space). [Figure 4-18] Figure 4: Alignment of orthologous proteins with the wild-type sequences of the SEQ ID NOs listed in Table 2 (CLUSTAL multiple sequence alignment with MUSCLE (3.8)). The motifs from Table 1 are highlighted in the alignment. The following symbols are used in the alignment: * -- all residues in that column are identical: -- conservative substitutions observed. -- semi-conservative substitutions observed. -- no match (space). [Figure 4-19] Figure 4: Alignment of orthologous proteins with the wild-type sequences of the SEQ ID NOs listed in Table 2 (CLUSTAL multiple sequence alignment with MUSCLE (3.8)). The motifs from Table 1 are highlighted in the alignment. The following symbols are used in the alignment: * -- all residues in that column are identical: -- conservative substitutions observed. -- semi-conservative substitutions observed. -- no match (space). [Figure 4-20] Figure 4: Alignment of orthologous proteins with the wild-type sequences of the SEQ ID NOs listed in Table 2 (CLUSTAL multiple sequence alignment with MUSCLE (3.8)). The motifs from Table 1 are highlighted in the alignment. The following symbols are used in the alignment: * -- all residues in that column are identical: -- conservative substitutions observed. -- semi-conservative substitutions observed. -- no match (space). [Figure 5] Figure 5: Example of the scale used for the assessment of damage-induced surface discoloration on leaf discs, represented by a description of each score and a photograph of a leaf disc with a given score. [Figure 6-1]Figure 6: Leaf discs from the phenotypic test of the F2 lane segregation analysis of lettuce plants containing mutation 1 and mutation 3, as described in Example 7. The photographs represent leaf discs after 1, 2, 3, 4 and 7 days of incubation after harvesting. [Figure 6-2] Figure 6: Leaf discs from the phenotypic test of the F2 lane segregation analysis of lettuce plants containing mutation 1 and mutation 3, as described in Example 7. The photographs represent leaf discs after 1, 2, 3, 4 and 7 days of incubation after harvesting. [Figure 7-1] FIG. 7: Phenotypic analysis of whole lettuce heads containing mutation 1 and mutation 3, as described in Example 8. [Figure 7-2] FIG. 7: Phenotypic analysis of whole lettuce heads containing mutation 1 and mutation 3, as described in Example 8. [Figure 7-3] FIG. 7: Phenotypic analysis of whole lettuce heads containing mutation 1 and mutation 3, as described in Example 8. DETAILED DESCRIPTION OF THE INVENTION

[0197] table [Table 2] [Table 3] [Table 4] [Table 5] [Table 6]

[0198] [Table 7]

[0199] Table 3 shows the F5H1 and F5H2 mutations and their effect on the encoded F5H1 and F5H2 protein sequences, with positions in the lettuce (Lactuca sativa) sequences SEQ ID NO: 115 (F5H1 CDS wild-type sequence), SEQ ID NO: 116 (F5H2 CDS wild-type sequence), SEQ ID NO: 1 (F5H1 protein wild-type sequence), and SEQ ID NO: 2 (F5H2 protein wild-type sequence). [Example]

[0200] Example Example 1 Generation of lettuce (Lactuca sativa) plant mutants by treatment with EMS To generate mutant lettuce plants, approximately 2,000 seeds of lettuce cultivars Troubadour, Apache, Yorvik, and Roderick were treated with EMS by soaking the seeds in an aerated solution of either 0.05% (w / v) or 0.07% (w / v) EMS for 24 hours at room temperature. After EMS treatment, the M seeds were rinsed with water, harvested, and germinated and grown in a greenhouse at 20°C under a 16-hour light / 8-hour dark period to generate bulked M seeds. The resulting population of M plants was screened for lettuce plants that exhibited reduced wound-induced discoloration using the phenotypic tests described in Example 2.

[0201] Lettuce plants that were mutants of cv. Troubadour and showed reduced wound-induced surface discoloration compared to the WT after 3 to 5 days of incubation of leaf disc samples in the phenotypic tests described in Example 2 were selected and crossed with a second lettuce plant derived from cv. Troubadour. An F6 line, 11K200310, was produced from this cross after repeated cycles of inbreeding, combining plant and line selection. Selection was performed to maintain the reduced wound-induced surface discoloration and reduce the effects of undesirable background mutations.

[0202] Five thousand seeds of line 11K200310 were then subjected to a second EMS treatment in a manner similar to that described above for the first EMS treatment, except that EMS was used at a concentration of 0.14%. The EMS-treated seeds were then sown in a greenhouse, and the resulting plant population was then screened again for lettuce plants with reduced wound-induced surface discoloration using the phenotypic test described in Example 2.

[0203] Example 2 Phenotypic identification of plants exhibiting reduced wound-induced surface discoloration Plants of Example 1 grown from seeds treated with either one or two EMS treatments were screened for their ability to exhibit reduced wound-induced surface discoloration. When approximately six true leaves had grown to a size that allowed for sample collection without cutting the middle vein of the leaf, leaf disk samples were collected from the plants. For this experiment, plants were grown in a greenhouse to a mature stage (approximately 2 months old). The obtained samples were placed on filter paper moistened with MES buffer. The upper side of the leaf was in contact with the filter paper. The leaf was covered with a second filter paper moistened with MES buffer. Air bubbles between the two filter papers were removed, and the leaf disks were incubated between the moist filter papers in a container at 7.5°C. After 3, 5, and 10 days of incubation of the leaf samples, wound-induced surface discoloration was independently scored for the presence and intensity of wound-induced surface discoloration, indicated by pink color around the edge of the leaf sample. An example of a scale used to distinguish this phenotype is 9 to 0, where 9 means no discoloration at the edge of the leaf disc, a score of 8 means the leaf disc has very slight pink discoloration around the edge, a score of 5 means the leaf disc has a thin ring of red / pink discoloration around the edge, a score of 2 means the leaf disc has a darker and thicker ring of red / pink discoloration around the edge compared to leaf discs with scores of 9, 8, 7, 6, 5, 4, or 3, and a score of 0 means the leaf disc has a significantly darker and thicker ring of red / pink discoloration around the edge. Different scales can be used to assess the intensity of wound-induced surface discoloration, but the scale should range from no wound-induced surface discoloration to maximum intensity of wound-induced surface discoloration. The most interesting candidates were grown, re-sown, and retested according to the same protocol.

[0204] Several candidates were confirmed to have reduced wound-induced discoloration. The results of phenotypic testing of wound-induced surface discoloration on leaf discs 3, 7, and 10 days after sampling are shown in Figures 1, 2, and 3, and the scores are shown in Table 4. On day 3 (Figure 1), WT plants showed wound-induced surface discoloration (score 0), plants containing Mutation 1 showed reduced wound-induced surface discoloration compared to that of wild-type plants (score 7), and plants containing Mutation 1 and Mutations 2, 3, 4, 5, or 6 did not show wound-induced surface discoloration (score 9). On day 5 (Figure 2), WT plants showed wound-induced surface discoloration (score 0), plants treated once with EMS and harboring mutation 1 in their F5H1 gene homolog (Table 3) showed reduced wound-induced surface discoloration compared to WT (score 0) (score 2), and plants treated twice with EMS and harboring mutation 1 in their F5H1 gene homolog (Table 3) and mutations 2, 3, 4, 5, or 6 in their F5H2 gene homolog (Table 3) showed no or reduced wound-induced surface discoloration compared to plants containing mutation 1 (scores 7–9). At day 10 (Figure 3), plants treated once with EMS and carrying mutation 1 in their F5H1 gene homolog showed wound-induced surface discoloration (score 0), whereas plants treated twice with EMS and carrying mutation 1 in their F5H1 gene homolog (Table 3) and mutations 2, 3, 4, 5, or 6 in their F5H2 gene homolog (Table 3) showed no or reduced wound-induced surface discoloration compared with plants containing mutation 1 (scores 5–9).

[0205] These plants were self-crossed to produce seeds which were then deposited at NCIMB under accession numbers NCIMB42546, NCIMB42547, NCIMB42548, NCIMB42549, NCIMB42550 and NCIMB42551.

[0206] [Table 8]

[0207] Example 3 Modification of lettuce F5H homologue DNA from lettuce plants treated with EMS and showing reduced wound-induced surface discoloration was analyzed to identify mutations in the F5H gene homologs using standard DNA sequencing techniques. A number of mutations in the gene homologs F5H1 and F5H2 in lettuce plants were identified.

[0208] A mutation in the lettuce F5H gene homolog on chromosome 4, designated herein as F5H1, resulted in a premature stop codon in the corresponding wild-type protein sequence shown in SEQ ID NO:1. A mutation in the lettuce F5H gene on chromosome 3, designated herein as F5H2, resulted in an amino acid change in the corresponding wild-type protein sequence shown in SEQ ID NO:2. The presence of the modified F5H1 protein in lettuce plants results in reduced wound-induced surface discoloration. The presence of modified F5H1 and F5H2 proteins in lettuce plants enhances this effect. The mutations shown in Tables 5 and 6 below were identified in the F5H gene homolog in lettuce plants. The tables show only a portion of the F5H1 and F5H2 sequences, including the mutated (substituted) nucleotide (SNP) and the adjacent 50 nucleotides on either side.

[0209] [Table 9]

[0210] [Table 10] [Table 11]

[0211] Example 4 Identification of F5H gene orthologues and conserved regions To compare the DNA and protein sequences of F5H1 and F5H2 from lettuce plants with those from other crop species, F5H gene orthologs in other crop species were identified using the BLASTN and BLASTP programs. The best hits per species were identified as candidate F5H1 and F5H2 gene orthologs. A non-exhaustive list of plants carrying one or more F5H gene orthologs is shown in the table below.

[0212] [Table 12]

[0213] The alignment revealed the presence of highly conserved amino acids among F5H gene orthologues from different species. Examples of highly conserved amino acid regions highlighted within the sequences of F5H gene orthologues in the protein sequence alignment (Figure 4) are shown in Table 1.

[0214] Example 5 Development of novel plants having the traits of the present invention A lettuce plant numbered 15E238260, which showed reduced wound-induced discolouration after 10 days of incubation and was found by the phenotypic test described in Example 2, was self-crossed. Using the DNA marker test based on SNP markers in Example 3, plant 15E238260 was shown to be homozygous for mutations in the F5H1 gene homolog and the F5H2 gene homolog. Flowers of plant 15E238260 were used as pollen donors to cross with lettuce cultivar Hofnar, which, like wild-type plants, showed reduced wound-induced surface discolouration in the phenotypic test described in Example 2. As a result of the above cross, an F1 seed lot numbered 15E97481 was obtained.

[0215] Four F1 seeds were sown, and the resulting plants, numbered 15E748101, 15E748102, 15E748103, and 15E748104, were self-crossed to produce F2 seeds. These F2 seeds were sown, and individual F2 plants were tested for reduced wound-induced surface discoloration by the phenotypic tests described in Example 2. As described in Example 2, three out of every 16 F2 plants were expected to show reduced wound-induced surface discoloration after 3-5 days of leaf disc incubation at 7.5°C, and one out of every 16 F2 plants was expected to show reduced wound-induced surface discoloration after 10 days of leaf disc incubation at 7.5°C.

[0216] Therefore, F2 plants that showed reduced wound-induced surface discoloration after 10 days of incubation were expected to be homozygous for the mutation in the F5H1 gene homolog and the mutation in the F5H2 gene homolog. The selected F2 plants were sown to produce F3 seeds. Ten F3 plants were grown from this seed lot, and the presence of homozygosity for the two mutant genes in the selected F2 plants was confirmed by observing reduced wound-induced surface discoloration for each of them after 10 days of incubation (as described in Example 2). Plant selection and confirmation of the selected genotype were performed using molecular markers that recognize the difference between the wild-type and mutant genes.

[0217] Example 6 Introgression of the traits of the present invention by backcrossing To introgress both the F5H1 and F5H2 gene homolog mutations into the lettuce cultivar Hofnar, backcrossing was performed using cultivar Hofnar as the recurrent parent. For this purpose, selected F2 plants that showed reduced wound-induced surface discoloration after 10 days of incubation by the phenotypic test described in Example 2 were used as parents in a cross with Hofnar. The resulting BC1 seeds were sown, and the BC1 plants were used as parents in a cross with Hofnar to produce BC2 seeds. Twenty BC2 seeds were sown, each of which was allowed to self-seed to produce BC2.S1 seeds. These 20 BC2.S1 seed lots (i.e., BC2.S1 families) were selected by sowing 20 BC2.S1 plants per seed lot, which were then segregated for both the F5H1 and F5H2 gene homolog mutations. For this purpose, 20 plants per BC2.S1 family were tested in the phenotypic test described in Example 2 after a 10-day incubation period. After these 10 days, BC2.S1 families that showed segregation for the reduced wound-induced discoloration phenotype were selected. From these families, BC2.S1 plants with reduced wound-induced surface discoloration phenotypes were selected and used as parents in a BC3 cross with Hofnar. These BC2.S1 plants were self-crossed to produce BC2.S2 seeds. Ten BC2.S2 seeds were grown into plants and tested in the phenotypic test described in Example 2 to confirm reduced wound-induced discoloration in all of them. After 10 days, they indeed showed no or reduced wound-induced discoloration in the phenotypic test. This confirmed the homozygous presence of mutations in the F5H1 gene homolog and the F5H2 gene homolog. Plant selection and confirmation of the selected genotype could also be achieved by using molecular markers that recognize the difference between the wild-type and mutant genes.

[0218] The resulting BC3 seeds were sown, and the BC3 plants were used as parents in crosses with Hofnar to produce BC4 seeds. 20 BC4 seeds were sown, and each of them was self-crossed to produce BC4.S1 seeds. These 20 BC4.S1 seed lots (i.e., BC4.S1 families) were sown, and 20 BC4.S1 plants per seed lot were sown to select BC4.S1 families, which were segregated for both F5H1 and F5H2 gene homolog mutations. To this end, each of the 20 plants per BC4.S1 family was tested in the phenotypic test described in Example 2 for a 10-day incubation period. After these 10 days, BC4.S1 families that showed segregation for reduced wound-induced surface discoloration phenotypes were selected. From such families, BC4.S1 plants with reduced wound-induced surface discoloration phenotypes were selected and could be used as parents in crosses of BC5 with Hofnar. The BC4.S1 plants were self-crossed to produce BC4.S2 seeds, and 10 of these BC2.S2 seeds were sown, all of which confirmed reduced wound-induced discoloration. Plant selection and confirmation of the selected genotypes could also be performed using molecular markers that recognize the differences between the wild-type and mutant genes, such as the SNP-based markers described in Example 3. The genotype of each selected plant could also be confirmed using molecular markers.

[0219] BC4.S2 seeds are sown in trials and the resulting plants are compared to plants of the cultivar Hofnar grown in the same trials, thus establishing whether the BC4 generation is sufficiently similar to Hofnar to be used by growers in practice.

[0220] Example 7 Segregation analysis of the trait of the present invention showed reduced damage-induced discoloration and mutation 1 (C in the F5H1 gene homolog 370 →T 370 ) and mutation 3 (G in the F5H2 gene homolog 494 →A 494Lettuce plants numbered 15E238260 containing the 15E238260 gene in a homozygous state were used for segregation analysis. Flowers of plant 15E238260 were used as pollen donors to cross with plants of the lettuce cultivar Troubadour, which, like wild-type plants, showed reduced wound-induced surface discoloration in the phenotypic tests described in Example 2, to obtain F1 seeds.

[0221] F1 plants grown from the F1 seeds were sown and self-crossed to obtain F2 seeds, which were used to analyze the segregation of the trait of the present invention.

[0222] The mutations were homozygously absent, heterozygously present, or homozygously present in the plants. In the present invention, the various genotypes and their designations are shown in Table 8. Reduced wound-induced surface discoloration was evaluated using the leaf disc test described in Example 2, except that for segregation analysis, leaf discs were taken from young plants (approximately 2 weeks old). The score for each leaf disc is shown in Table 9, and an example of a leaf disc from a lettuce plant containing mutations 1 and 3 is shown in Figure 6.

[0223] [Table 13]

[0224] [Table 14]

[0225] The results in Table 9 show that to show reduced wound-induced surface discoloration, plants required a mutation in the F5H1 gene homolog (C 370 →T 370) is required to carry mutation 1. Preferably, the plants homozygously carried mutation 1. Plants homozygously carrying mutation 1 (BB) and heterozygously (AB) or homozygously (BB) carrying mutation 3 showed reduced wound-induced surface discoloration compared to wild-type (AA / AA) and plants carrying mutation 1 alone. The greatest reduction in wound-induced surface discoloration was observed in plants homozygously carrying mutation 1 in the F5H1 gene and homozygously carrying mutation 3 in the F5H2 gene (BB / BB).

[0226] Example 8 Phenotypic analysis of a single head of lettuce Lettuce plants were harvested at maturity (approximately 3 months old), and preferably the central (average) plant of the plot was selected for phenotypic analysis. The harvested plants were stored overnight in a cold room at 5°C in a plastic-wrapped box to prevent drying. The plants were then cut at low temperature (approximately 15°C). Old leaves, leaves showing blight symptoms, and outer leaves were removed from the heads. Heads of plants containing Mutation 1, Mutation 1 and Mutation 2, Mutation 1 and Mutation 3, Mutation 1 and Mutation 4, Mutation 1 and Mutation 5, and Mutation 1 and Mutation 6, as well as two or three heads of wild-type plants (Troubadour), were cut vertically into four sections to remove the cores, and the four sections were further cut horizontally two or three times. For phenotypic analysis, 100 g of cut lettuce leaves were washed in a washing machine (bubble washing step: 3 minutes, centrifugation time: 2.5 minutes at maximum speed), with the water replaced after each washing step. For small-scale washing, the water should be as cold as possible and the washing should be done in a large sink. The cut and washed leaves are dried in a salad spinner.

[0227] The cut leaves were folded twice and placed in ethylene-free dry plastic bags, which were then placed in boxes and stored at 5-6° C. The bags were not allowed to overlap each other.

[0228] An example of phenotypic analysis of the entire lettuce head is shown below. The wild-type lettuce cultivar Troubadour (wild type) does not contain the modified F5H gene homolog, the mutant 1 (C370 →T 370 ) (“Mutation 1”) and lettuce plants containing mutations 1 and 3 (G in the F5H2 gene homolog 494 →A 494 ) ("Mutation 1 and Mutation 3") are shown in FIG. 7. Photographs were taken 7 and 10 days after washing. Wild-type plants showed wound-induced surface discoloration at the cut edge 7 days after washing, plants containing Mutation 1 showed reduced wound-induced surface discoloration at the cut edge 7 or 10 days after washing compared to the wild type, and plants containing Mutation 1 and Mutation 3 showed no wound-induced surface discoloration even 10 days after washing, and therefore reduced wound-induced surface discoloration compared to plants containing Mutation 1.

[0229] Evaluation of wound-induced surface discoloration 3, 7, and 14 days after washing was performed using a separate assay. The results are shown in Table 10. Plants containing mutation 1 showed reduced wound-induced surface discoloration compared to the wild-type. Plants containing mutation 1 and mutations 3, 4, 5, or 6 showed reduced wound-induced surface discoloration compared to the wild-type plants and plants containing mutation 1. Plants containing mutation 1 and mutation 2 showed the same wound-induced surface discoloration as plants containing mutation 1 at day 14, but the wound-induced surface discoloration was slower than that of plants containing mutation 1, with plants containing mutation 1 having a score of 7 at day 7 and plants containing mutations 1 and 2 having a score of 8.

[0230] [Table 15]

[0231] array protein SEQ ID NO: 1 >Lettuce_LsF5H_1_1 MESLQIPIAFYAIIAILTFFFLSWVRRKPLPPGPMGWPIIGNMLMMDQLTHRGLARLAEKYGGILHLKMGFSHTIAVSSPEMARIILQEKDNIFANRPATIAITYLTYNGVDLAFANYGPFWRQMR KLCVMKLFSRKRAESWDSVRDEVDTMVKATAINSGTPVNLGELVFGLTHDIIYRAAFGSISHEGKEEFIRILQEYTKLFGAFNLADFIPFLGFIDPAGLNTRLPAARAALDGFIDKIIDEHLRKGKK TGDEGLDNDMVDEMLAFYSEEGKVNEGGDLQNAINLTRDNIKAIIMDVMFGGTETVASAIEWAMTELMHTPEALKRVQQEMANVVGLDRRVEESDLEKLTYFKCVIKETLRLHPPIPVLLHQSSEA TEVSGYHIPKGTRVMVNAYAINRDKNSWEDPDTFNPSRFLQNGAPDFRGSNYEFLPFGSGRRSCPGMQLGLYAMEVAHLLHCFTWELPDGMKPSEIDMGDVFGLTAPKAIRLVAVPTPRLLCPLY SEQ ID NO: 2 >Lettuce_LsF5H_2_1 MDPKSILLYVVLPLLTFFLLSRLRRKPLPPGPRGWPLIGNMLMMDQLTHRGLARLGEKYGGLLHLKMGFSHTVAVSSPEIARQVLQVQDNIFANRPATIAISYLTYDRQDMAFANYGPFWRQMRKLCVMKLFSRKRAESWDSVRDEVVSMVKITAASSGTAVNLGELVFGLTHDIIYRAAFGSISHEGKEEFIRILQEYTKLFGAFNLADFVPWLGFIDPAGLNTRLPCARAALDRIFKIIDEHLAKERK TGDEEDNDMVDEMLAFYSEEGKVNEGEDLQNAIRLTRNNIKAIIMDVMFGGTETVASAIEWALTELMHTPESLKRAQQELADVVGLDRRVEESDFEKLTYFKCVIKETLRLHPPIPVLLHQSSEATSVAGYHIPKGTRVMVNAFAINRDKNSWKDPHTFNPSRFLQDGAPDFKGSNYEFLPFGSGRRSCPGMQLGLYAMEVAHLLHSFTWQLPDGMKPSEIDMNDVFGLTAPKAIRLVAVPTPRLLCPLY sequence number 3 >Endive_ce_gene1 MDYLQIPIPIYAIIAILTFFFLAWLRRKPLPPGPMGWPIIGNMLMMDQLTHRGLASLANKYGGILHLKMGFSHTIAVSSPEIARQILQEKDNIFANRPATIAITYLYTYNRVDMAFANYGPFWRQMRKLCVMKLFSRKRAESWDSVRDEVDTMVKATATNSGLPVNLGELVFGLTHDIIYRAAFGSISHEGKEEFIRILQEYTKLFGAFNLADFIPMLGFIDPAGLNTRLPAARAALDGFIDKIIDEHLSKEKK TGDENVDNDMVDEMLAFYSEDGRINEGGDLQNAINLTRDNIKAIIMDVMFGGTETVASAIEWTMTELMHTPEALXRVQQELTNVVGLDRRVEESDFEKLTYFKCVIKETLRMHPPIPVLLHQSSEATEVSGYHIPKGTRVMVNAYAINRDKXSWEDPDTFNPSRFLQNGAPDFRGSNYEFLPFGSGRRSCPGMQLGLYAMEMAVAHLLHCFTWDLPDGMPSEIDMGDVFGLTAPKAIRLVAVPTPRLLCPLY sequence number 4 >Endive_ce_gene2 MDSLQIPIPVYAIIAILTFFFLAWLRRKPLPPGPIGWPIIGNMLMMDQLTHRGLASLAKKYGGILHLKMGFSHTIAVSSPEIARQILQEKDNIFANRPATIAITYLYTYNRVDMAFANYGPFWRQMRKLCVMKLFSRKRAESWDSVRDEVDTMVKATAINSGLPVNLGELVFGLTHDIIYRAAFGSISHEGKEEFIRILQEYTKLFGAFNLADFIPMLGFIDPAGLNTRLPAARAALDGFIDKINIDEHLSKEKK TGDENVDNDMVDEMLAFYSEDGRVNEGGDLQNAINLTRDNIKAIIMDVMFGGTETVASAIEWTMTELMHTPEALKRVQQELTNVVGLDRRVEESDFEKLTYFKCVIKETLRMHPPIPVLLHQSSEATEVSGYHIPKGTRVMVNAYAINRDKNSWEDPDTFNPSRFLQNGAPDFRGSNYEFLPFGSGRRSCPGMQLGLYAMEMAVAHLLLCFTWELPDGMKPSEIDMGDVFGLTAPKAIRLVAVPTPRLLCPLY sequence number 5 >Endive_ce_kethel_v0.1_EVM71979 MDPMSILLYVVLPLFTFFLLSRFRRKPLPPGPRGWPVIGNMLMMDQLTHRGLARLGEKYGGLLHLKMGFSHTVAVSSPEIARQVLQVQDNIFANRPATIAISYLTYDRQDMAFANYGPFWRQMRKLCVMKLFSRKRAESWDSVRDEVVSMIKITAASSGSAVNLGELVFGLTHDIIYRAAFGSISHEGKEEFIRILQEYTKLFGAFNLADFIPWLGFIDPAGLNNRLPKARAALDGFIDKIIDEHLRKEKK TGDEEDNDMVDEMLAFYSEEGKVNEGEDLQNAIRLTRNNIKAIIMDVMFGGTETVASAIEWALTELMHTPESLKRAQQELVDVVGLDRRVEESDFEKLTYFRCVIKETLRLHPPIPVLLHQSSEATEVAGYHIPKGTRVMVNAFAINRDKNSWKDPHTFNPSRFLQDGAPDFKGSNYEFLPFGSGRRSCPGMQLGLYAMEMAVAHLLHSFTWQLPDGMKPSEIDMSDVFGLTAPKAIRLVAVPTPRLLCPLY sequence number 6 >Chicory_ci_vitessa_fr_v1_EVM170737 MDSLQIPIPVYAIIAVLTFVFVAWLRRKPLPPGPMGWPIIGNMLMMDQLTHRGLASLAKKYGGILHLKMGFSHTIAVSSPEIARQILQEKDNIFANRPATIAITYLYTYNRVDMAFANYGPFWRQMRKLCVMKLFSRKRAESWDSVRDEVDTMVKATATNSGLPVNLGELVFGLTHDIIYRAAFGSISHEGKEEFIRILQEYTKLFGAFNLADFIPMLGFIDPAGLNTRLPAARAALDGFIDKIIDEHLSKEKK TGDENVDNDMVDEMLAFYSEDGRINEGGDLQNAINLTRDNIKAIIMDVMFGGTETVASAIEWTMTELMHTPEALKRVQQELTNVVGLDRRVEESDFEKLTYFKCVIKETLRMHPPIPVLLHQSSEATEVSGYHIPKGTRVMVNAYAINRDKNSWEDPDTFNPSRFLQNGAPDFRGSNYEFLPFGSGRRSCPGMQLGLYAMEMAVAHLLHCFTWELPDGMKPSEIDMGDVFGLTAPKAIRLVAVPTPRLLCPLY sequence number 7 >Chicory_ci_vitessa_fr_v1_EVM170780 MDSLQIPIPVYAIIAILTFFFLAWLRRKPLPPGPMGWPIIGNMLMMDQLTHRGLASLAKKYGGILHLKMGFSHTIAVSSPEIARQILQEKDNIFANRPATIAITYLYTYNRVDMAFANYGPFWRQMRKLCVMKLFSRKRAESWDSVRDEVDTMVKATAINSGLPVNLGELVFGLTHDIIYRAAFGSISHEGKEEFIRILQEYTKLFGAFNLADFIPMLGFIDPAGLNTRLPAARAALDGFIDKINIDEHLSKEKK TGDENVDNDMVDEMLAFYSEDGRVNEGGDLQNAINLTRDNIKAIIMDVMFGGTETVASAIEWTMTELMHTPEALKRVQQELTNVVGLDRRVEESDFEKLTYFKCVIKETLRMHPPIPVLLHQSSEATEVSGYHIPKGTRVMVNAYAINRDKNSWEDPDTFNPSRFLQNGAPDFRGSNYEFLPFGSGRRSCPGMQLGLYAMEMAVAHLLHCFTWELPDGMKPSEIDMGDVFGLTAPKAIRLVAVPTPRLLCPLY sequence number 8 >Chicory_ci_vitessa_fr_v1_EVM158591 MDPMSILLYVVLPLFTFLLSRLRKRKPLPPGPRGWPVIGNMLMMDQLTHRGLARLGEKYGGLLHLKMGFSHTVAVSSPEIARQVLQVQDNIFANRPATIAISYLTYDRQDMAFANYGPFWRQMRKLCVMKLFSRKRAEWDSVRDEVISMIKITAASSGSAVNLGELVFGLTHDIIYRAAFGSISHEGKEEFIRILQEYTKLFGAFNLADFIPWLGFIDPAGLNTRLPKARAALDGFIDKIIDEHLSKEKKTGDEEDNDMVDEMLAFYSEEKGVNEGEDLQNAIRLTRNNIKAIIMDVMFGGTETVASAIEWALTELMHTPESLKRAQQELADVVGLDRRVEESDFEKLTYFRCVIKETLRHPPIPVLLHQSSEATEVAGYHIPKGTRVMVNAFAINRDKNSWKDPHMFNPSRFLQDGAPDFKGSNYEFLPFGSGRRSCPGMQLGLYAMEMAVAHLLHSFTWQLPDGMKPSEIDMSDVFGLTAPKAIRLVAVPTPRLLCPLY sequence number 11 >Celery_RZ_draft_99.605_EVM363634 MSLLFILLLTIISMIFFISRFGGKPYPPGPRRLPVIGNTLMMACLTHRGLAKLASRYGGLFYLRMGAQDIVVVSTPDMARQILQTHQDQISNRPTSIALDYLSYGRANMAFADYSPWWRQMRKICVMKLFSRARVESWNSVRDELNHMLRDVASNANQAINLGELVFGHTEKIIYRAAFGSRLSDGGTEFIKIMQEFSKLFGSFNVCDFVPWMSRADPQGLKARLRKARGSLDTFIDSIIDQHIIKRKGKN IGDKDMVDELLAFYTEEGEAKAESDDLQATIKLTNIKGIIMDIMFGGTETVAAAIEWAMSELKNPEELRKTQEELSNVVGLHRCVEEGDLEKLTYLKCVLKETLRHLHPPLPFLFPRAAEDVYVAGYYIPAGSRVIINLWAMGHDGKCWNDEPEAFKPSRFLDVGAPDYRNNNFEFIPFGTGRRSCPGMQLGLHAFEMGLAHLLHCFNWELPDGMKPSQVDMMYGLSAPKATRLIAVPTPRLLCPIC sequence number 12 >Celery_RZ_draft_99.605_EVM348724 MTTIFFLLLLCLFLARRKPYPPGPKGWPIIGNMLIMDKLTHRGLAKIATQYGGIVHLRMGFLHMVTVSTPDMVREVLQIQDTVFANRPATMNISYLTYNRADMAFANYGPWRQMRKISIIKLFSRKRAESWDSVRDQVDDMLGKVVSNSGLSVNIGELVFGLTRNIIYRAAFGSISGQGQDEFIKIMQEFSKLFGAFNICDFVPGITWLDPQGFKVRLVKARESLDKFIDSILDEHIANQKSNLNGST DEGNGDMVYQLLAFYSEEQSKVNHSDDTNNALKLTRDNIKAIIMDVMFGGTETVASAIEWAMSELMGSPEDLKRVQQELIDVVGLHRRVEENDFDKLIYLKCCIKETLRLHPPIPLLLHETAQEAVVAGYHIPAKSRVMINSWAVNRDPNSWDDPEKFKPSRFLKQGMPDFKGSNFEFIPFGSGRRSCPGMQLGLYAFEMAVAHLLHSFNWELPDGMPSELDTNDVFGLTAPRATRLVAVPTPRLLCSIC sequence number 13 >Celery_c17480_g1_i1|m.26831 METNTTAMTILFFILPLLSFFLLSSFRRKRYPPGPKGWPIIGNLLMMDKLSHRGLAKLAAQYGGLVHLRMGFLHMFTVSTPDMAREVLQIQDNIFANRPATMNISYLTYDRADMAFANYGPFWRQMR KISVMKLFSRKRAESWDSVREEVDDMVKIVLSKTGCSVNIGELVFGLTRNIIYRAAFGTLSHEGQDEFIKILQEFSKLFGAFNICDFVPGLTWADPQGFMGRVVKARASLDGFIDSIIDAHIEKKKSS KNGIIDEGNSDMVYELLDFYGEEKAKVSEFEDQNSSLKLTRDNIKAIIMDVMFGGTETVASAIEWAMSELMRSPKDLKKVQQELVNVVGLHRRVEESDFDKLTYLKCCIKETLRLHPPIPLLLHETAQ DAEVAGYHIPARSRVIINSWAINRDPNSWTDPDTFKPSRFLQEGMPDFKGSNFEFIPFGSGRRSCPGMQLGLYALEIAVAHLLHCFNWELPDGMKPSEVDTDDVFGLTAPRATRLVAVPTPRLLCPIS SEQ ID NO: 14 >Brassica_Bo1g005770 MESSISQTLSQVLDPTTGILIVVSLFIFIGLITRGRRPPYPPGPRGWPIIGNMSMMDQLTHRGLANLAKKYGGLCHLRMGFLHMYAVSSPDVARQVLQVQDSVFSNRPATHIAISYLTYDRADMAFAHYGPFWRQMRKVCVMKVFSRKRAESWASVRDEVDRMIRSVSSNVGKSINVGEQIFALTRNITYRAAFGSACEKGQDEFIRILQEFSKLFGAFNVADFIPYFGWIDPQGINKRLVKARNDLDGFIDDIIDEHIKK KENQNSIDAGDVVDTDMVDDLLAFYSEEAKLVSETADLQNSIKLTRDNIKAIIMDVMFGGTETVASAIEWALTELLRSPEDLKRVQQELAEVVGLDRRVEESDIEKLTFLKCTLKETLRLHPPIPLLLHE TAEDTEIDGYFVPKKSRVMINAFAIGRDKNSWVDPETFRPSRFLEPGVPDFKGSNFEFIPFGSGRRSCPGMQLGLYALELAVAHILHCFTWKLPDGMKPSELDMSDVFGLTAPKATRLYAVPSTRLICSV sequence number 15 >Brassica_Bo7g117840 MESSVSQTLSQVLDPTTAILIVVSLFIFIGLITRRRRSYPPGPRGWPIIGNMLMMDQLTHRGLANLAKKYGGLCHLRMGFLHMYAVSSPDVARQVLQVQDSIFSNRPATIAISYLTYDRADMAFAHYGPFWRQMRKVCVMKVFSRKRAESWASVRDEVDKMIRSVSSNVGKSINVGEQIFALTRNITYRAAFGSACEKGQDEFIRILQEFSKLFGAFNVADFIPYFGWIDPQGINKRLVKARNDLDGFIDDIIDEHMKKKENQNTVDDGYVGDTDMVDDLLAFYSEEAKLVSETTDLQNSIKLTRDNIKAIIMDVMFGGTETVASAIEWALTELLRSPEDLKRVQQELAEVVGLDRRVEESDIEKLTFLKCTLKETLRHPPIPLLLHETAEDTEIDGYFVPKRSRVMINAFAIGRDPKSWPDAETFRPSRFLEPGVADFKGSNFEFIPFGSGRRSCPGMQLGLYALELAVAHILHCFTWKLPDGMKPSELDMNDVFGLTAPKATRLFAVPSTRLICAV sequence number 16 >Brassica_Bo7g119430 MDCLLCSPILYVVLIFLWYLVRVLLTRSNPFPPGPKGYPIGNMKLKNQLNHRGLAELAKQYGGLLHLQMGRIHIVAASTAEMAREILQVQDVVFANRPNVAISYLTYNRADFANYGPLWRQMRKVCVMKLFSRKRAESWASVRDEINTMVQTLTKQTGSPVNVGELVFALTRNITYRAAFGSFARDGQDEFVKILQEFSKLFGADFLTPFWMKWFSNRDFSKRLENARKSLDGF IDRIIDAHIEKKNSRKQDDDGLEDDMVDELMAFYSGESGENGGKSNDSLSSSFKLTRDNIKALVMDVMFGGTETVASAIEWAMTELMKNPHELVKLQQELADVIGLNREFHESDLENLPYFRCAMKETLRHPPIPLLLHEAADSVVSGYSIPRDSRVMINVYAIGRDGSVWTEPDAFRPGRFMDSKAPDFKGSDFEFLPFGSGRRSCPGMQLGLYAMELAVAHMLHSFDWELPEGGSSDDDLDMTDMFGLTAPRATRLIAVPSYRLKCPMVI sequence number 17 >Brassica_Bo3g093960 MESLLSQTLNQVIDPTPSVLLITISLLVVVYLISQWFKPLYPPGPKGLPVIGNMLMMDQLTHHGLAKLAHKYGGLFHLRMGFRHVFAITSPDVARQVLQVQDISFSNRPVNVAINYLTYDLADMAFAPYGPFWREMRKVCVMKVFSRKRTESWASVREEVNNMVRSFSSNVGKPVNVGELIFTLTRNITYRAAFGAACETEQDEFIRILQEFSKLFGAFNIADFIPFLGWFDFQGINKRLVKARNDLDGFIDEVIDE HMKKRETVNVDEDTDMVDDLLAFYSEDSSTNRNKNTVKLTRDNIKALVMDVMFGGTETMASGIEWALTELLRNPAELKRLQQELTEVVGLDRRVDDTHLEQLTFLKCTLKETMRLHPPIPLILHEAIEDRKLQGFFVPKGSRLMINAFAIARDPKLWVDPEAFRPSRFMEPGMPDFMGTNFEFIPFGAGRRSCPGMQLGLYAMEVAVANIIHCFTWKLPDGMKPSELDMSDVMGLTPRATRLIAVPDTRLICPVYP sequence number 18 >Brassica_XP_013607401.1 MYTLMTLILLVPLLFLFRHLLSRRLRQRKPYPPGPKGLPIIGNILMMNQFNHRGLAKLSRTYGGLLHLLRGISHLFVVSSPQIARQVLQVQDHVFSNRPTTIAIRYLTYGQSDLAFCNYGPFWRRMRK LYVMMLFSRKRAESWASVDEEVHKAVRSVAANVGKPLNVCKVAFSLTRDITFRAAFGSSSSSNEGRLDEFLEIIQEFSKLFGEFNVADYVPSWLSWIDPQGINKRVEKARKSLDCFIESIINDHLDKK KTEKNVNVDEVTDMVDQLLAFYKEEVKVKDSETKINLDNIKGIIMDVMFGGTETVALAIEWVLTELLRSPENMKRVQDELATVVGLERWSVEDTHLEKLSFLKVCLVKDTLRLHPPFPLLLHETVEEAEV SGYFIPKGSRVMVXTYALGRDPASWSSDPEIFNPSRFLDPGAPDLKGNSFEFIPFGSGRRSCPGMQLGMYAFELAVAHLLHCFTWRLPDGVKFGDVDTIEGPGLTVAKANSLVAVPIKRLLCPMVLESHNV sequence number 19 >Eggplant_c18725_g1_i1|m.19489 MKEMVQKNINSILEALQANPMLLFLFIIPLFFLYLFSTSSRRYPPGPRGWPLIGNMMIMDQLTHRGLAKLAEKYGGVMHLKMGYIHKIVISGPEEARQVLQVQDNIYSNRPATVAISYLTYDRADMAFAD YGPFWRQMRKLCVMKLFSRKRAESWDSVRDEVDSLIKIVTTNAGTSVNLGELVFGLTRDIIYRAAFGTSSAEGQEEFIKILQEFSKLFGAFNMVDFIPWLGWIGQQGLNVRLANARASLDGFIDSIIDDHI ERKKANVTIDNGDRETDMVDELLAFYSEEATVNESEDNLQNAIRLTRDNIKAIIMDVMFGGTETVASAIEWVMAELVKNPEELKKVQQELANVVGLNRRVEESDLEKLTYLKCCLKETLRLHPPIPLLLHE TAEESTIFGYHIPAGSHVVINSFAIGRDKNSWEDADSYKPSRFLKQGVPDFKGGNFEFLPFGSGRRSCPGMQLGLYALEMAVHLLHCFTWELPDGMKPSELKMDDTFGLTAPLANRLVVVPSPRLLCTLY sequence no. 20 >Eggplant_sm_67_3_v1_EVM112823_1_ MKEIIQNNTFFILTTKETNLMMPLYFIFPIFIFFFFTISRLNKKNFPPGPKGWPIIGNMMMMDQLTHHGLAKLAKKYGGILHQMGYLQMTVVSSPAEAREVLQLQDTIFANRPTTIAVEYLSYARADMAFANYGPFWRQMRKLCVMKLFSRKRAESWDSIRDEVESMTMAVATRVGSSVNIGELVFGVAKNIIYRAAFGTCSNRGQDELLNIMQEFSKLFGAFNLADFIPWLGWVDPQGLNKMIIKARASLDGFIDTIIDDHIQRKKTNNYNDEGNNDMVDELLAFYGEKTKLNDSDDLTNALRLTRDNIKSIIMDIMFGATETVASAIEWAMAELMKSPEDLKMVQQELANVVGLHRKVEEMDFEKLIFLKCCIKETLRLPPIPLLVHESVEDTTINNYIIPAKSRVIVNAWAIGRDKNSWDDPESFKPSRFLKEGVADFKGGDFEFLPFGSGRRSCPGMQMGLYAFEMTLAHLLHCFNWELPNGMKPSDIDDMNDVFGLTVPKATRLVAPTPPRLLCQLY sequence no. 21 >Potato_XP_006340697.1 MKEMVLNNINSTLEALQAQPMLLFFFIIPLFLYLFSTSRRKRYPPGPLGWPLIGNMMMMDQLTHRGLAKLAQKYGGVFHLKMGYVHKIVISGPEEARQVLQVQDNIYSNRPKTVAISYLTYDRADMAFA DYGPFWRQMRKLCVMKLFSRKRAESWDSVRDEVDSMVKIVTNTGTSINLGELVFCLTRNIIYRAAFGTSSDEGQDDFIKILQEFSKLFGAFNMADFIPWLRWIGQQGLNVRLAKARASLDGFIDSIIDDH IERKKANVINDDGYRESDMVDELLAFYSEETKVNESEDLQNSIRLTRDNIKAIIMDVMFGGTETVASAIEWAMAELMKSPEDLKKVQQELANVVGLNRKVDESDFEKLTYLKCCLKETLRLHPPIPLLLH ETAEESTVSGYYIPAKSHVIINSFAIGRDKNSWEDPDSFKPSRFLKEGVPDFKGGNFEFLPFGSGRRSCPGMQLGLYALEMAVHLLHCFTWELPDGMKPSELKMDDIFGLTAPLANRLVAVPTPRLLCNY sequence no. 22 >Apple_XP_008372753.1 MDYLLQSLQPLQSMTPLLLIIPLLFLLPLIFRFRRPPPYPPGPKGLPLIGNMLLMDQLTHRGLAKLAKKYGGIFHLRMGFLHMVAISNPDVARQVLQVQDNIFSNRPATIAISYLTYDRADMAFAHYGP FWRQMRKLCVMKLFSRKRAESWESVRDEVDSSVRTVTVHVGSAVNIGELVFSLTKNIIYRAAFGTSSQEGQDEFIGILQEFSKLFGAFNIADFIPSLGWVDPQGLNNRLAKARESLDRFIDTIIDDHME KKRNGKGVSDGETDMVDELLAFYSEAKVYESEDNLQNAIKLTRDNIKAIIMDVMFGGTXTVASAIEWAMSELMKSPEDLKRVQQELADVVGLDRRPEETDFEKLTYLKCALKETLRLHPPIPLLLHETSEDAVVAGYRIPKRSRVMINAWAIGRDKDSWEDAESFKPSRFLKEGVPDFKGSNFEFIPFGSGRRSCPGMQLGLYALEMAVHLLHCFTWELPDGMKPSELDMNDVFGLTAPRASRLIAVPSKRVVCPL sequence no. 23 >Apple_XP_008337913.1 MDSLLQSLQPLKSMTPLVFIIPLLFLLPLIFCFRRPPYPPGPKGLPLFGNLMMDQLTHRGLAKLAKQYGGIFHLRMGFLHMVAISNPDVARQVLQVQDNIFSNRPATIAISYLTYDRADMAFAHYGP FWRQMRKLCVMKIFSRKRAESWESVRDEVDTAVRTVTVHVGSAVNIGELVFSLTKNIIYRAAFGTSSQEGQDEFIGILQEFSKLFGAFNIADFIPSLGWVDPQGLNNRLAKARESLDRFIDTIIDDHME KKKKNNKGLNDGETDMVDDLLAFYSEEAKVNESEDNLQNAIKLTRDNIKAIIMDVMFGGTETVASAIEWAMSELMKNPEDLKRVQQELLNVVGLDRRPEEADFEKLTYLKCALKETLRLHPPIPLLLHE TSEDAVVAGYHIPKKSRVMINAWAIGRDKDSWEDPESFKPSRFLKEGVPDFKGSNFEFIPFGSGRRSCPGMQLGLYALEMAVAHMLHCFTWELPDGMKPSELDMNDVFGLTAPRASRLVAVPSKRVVCPL sequence no. 24 >Apple_XP_008391587.1 MELLHQALQSLQSSPMLLILLLLLIISFIFLFNSRRKPPYPPGPKGWPIIGNMLMTDQLTHRGLAHLAKQYGGLLHLQMGVIHVMAVSTDPDMAREILQSQDSLFANRPANVAISYLTYDRADMAFANY GPFWRRMRKICVINLFSRKRAESWASVREEVDEMVQTVAGKTGSPVNIGQLVFALTRNITYRAAFGSSSHEGQGEFVQILQEFSKLFGAFNMQDFLPWLGWVHAQGFQDRMARARKSLDVFIDKIIDDHM AKRKANMEKDDSEAADTDMVDELIAYFSDDAGKEGDDPNSGFKLTRDNICALIMDVMFGGTETVASVIEWTMAELMKSPEDLKRVQQELTDVVGLNRRLQETDLENLTYLKCAVKESLRLHPPIPLLLHETVEDTSVAGYSFPAGSRVWINAWAIARDPTAWDEPETFKPSRFLEDSSPDFKGSNFEFIPFGSGRRSCPGMALGLEMAVAHLLHCFAWELPGGMKPSELDMNDVFGLTAPKAVQLVAVTYRLNCPL sequence no. 25 >Pear_XP_009378215.1 MDSLQSLQALQSMTPLLLIIPLLFLLPLIFRFRRPPPYPPGPKGLPLIGNMLLMDQLTHRGLAKLAKKYGGIFHLRMGFLHMVAISNPDVARQVLQVQDNIFSNRPATIAISYLTYDRADMAFAHYGP FWRQMRKLCVMKLFSRKRAESWESVRDEVDSAVRTVTVHVGSAVNIGELVFSLTKNIIYRAAFGTSSQEGQDEFIGILQEFSKLFGAFNIADFIPSLGWVDPQGLNNRLAKARESLDRFIDTIIDDHME KKRNGKGVSDSETDMVDELLAFYSEEAKVNESEDNLQSAIKLTRDNIKAIIMDVMFGGTETVASAIEWAMSELMKSPEDLKRVQQELADVVGLDRRPEETDFEKLTYLKCALKETLRLHPPIPLLLHETSEDAVVAGYRIPKRSRVMINAWAIGRDKDSWEDAESFKPSRFLKEGVPDFKGSNFEFIPFGSGRRSCPGMQLGLYALEMAVHLLHCFTWELPDGMKPSELDMNDVFGLTAPRASRLIAVPSKRVVCPL sequence no. 26 >Pear_XP_009338655.1 MDSLLQSLQPLQSMTPLLLIIPLLFLLPLIFRFRRPPPYPPGPKGLPLIGNMLLMDQLTHRGLAKLAKKYGGIFHLRMGFLHMVAISSPDVARQVLQVQDNIFSNRPATIAISYLTYDRADMAFAHYGP FWRQMRKLCVMKLFSRKRAESWESVRDEVDSAVRTVTVHVGSAVNIGELVFSLTKNIIYRAAFGTSSQEGQDEFIGILQEFSKLFGAFNIADFIPSLGWVDPQGLNNRLAKARESLDRFIDTIIDDHME KNRNGKGVSDSETDMVDELLAFYSEEAKVNESEDNLQSAIKLTRDNIKAIIMDVMFGGTETVASAIEWAMSELMKSPEDLKRVQQELADVGLDRRPEETDFEKLTYLKCALKETLRLHPPIPLLLHET SEDAVVAGYRIPKRSRVMINAWAIGRDKDSWEDAESFKPSRFLKEGVPDFKGSNFEFIPFGGSGRRSCPGMQLGLYALEMAVAHLLHCFTWELPDGMKPSELDMNDVFGLTAPRASRLIAVPSKRVVCPL SEQ ID NO: 27 >Pear_AGR44939.1 MDSLLQSLQPLKSMTPLVFIIPLLFLLPLIFRFRRLPPYPPGPKGLPLIGNMLMMDQLTHRGLAKLAKQYGGIFHLRMGFLHMVAVSNPDVARQVLQVQDNIFSNRPATIAISYLTYDRADMAFAYYGP FWRQMRKLCVMKLFSRKRAESWESVRDEVDSAVRTVTVHVGSAVNIGELVFSLTKNIIYRAAFGTSSQEGQDEFIAILQEFSKLFGAFNIADFIPSLGWVDPQGLNNRLAKARESLDRFIDTIIDDHME KKKNNKGLNDGETDMVDELLAFYSEEAKVNESEDNLQSAIKLTRDNIKAIIMDVMFGGTETVASAIEWAMSELMKSPEDLKRVQQELADVVGLDRRPEETDFEKLTYLKCALKETLRLRPPIPLLHET SEDAVVAGYRIPKRSRVMINAWAIGRDKDSWEDAESFKPSRFLKEGVPDFKGSNFEFIPFGSGRRSCPGMQLGLYALEMAVHLLHCFTWELPDGMKPSELDMNDVFGLTAPRASRLIAVPSKRVVCPL sequence no. 28 >Pear_XP_009346304.1 MDSLQSLQPFTSMTPLVFIIPLLFLLPLIFRFRRLPPYPPGPKGLPLIGNMLMMDQLTHRGLAKLAKQYGGIFHLRMGFLHMVAVSNPDVARQVLQVQDNIFSNRPATIAISYLTYDRADMAFAHYGP FWRQMRKLCVMKLFSRKRAESWESVRDEVDTAVRTVTVHVGSAVNIGELVFSLTKNIIYRAAFGTSSQEGQDEFIAILQEFSKLFGAFNIADFIPSLGWVDPQGLNNRLAKARESLDRFIDTIIDDHME KKKNNNKGLNDGETDMVDDLLAFYSEEAKVNESEDNLQNAIKLTRDNIKAIIMDVMFGGTETVASAIEWAMSELMKSPEDLKRVQQELFNVVGLDRRPEEADFEKLTYLKCALKETLRLHPPIPLLLHE TSEDAVVSGYHIPKQSRVMINAWAIGRDKDSWEDPESFKPSRFLKDGVPDFKGSNFEFIPFGSGRRSCPGMQLGLYALEMAVAHMLHCFTWELPDGMKPSELDMNDVFGLTAPRASRLVAVPSKRVVCPL sequence no. 29 >Pear_XP_009378429.1 MELLRQALQSLQSSPMLLILLLLLVISLIFLFNSRRKLPYPPGPKGWPIIGNMLMTDQLTHRGLAHLAKQYGGLLHLQMGAIHVIAVSTPDTAREILQVQDSSFANRPANIALSYLTYDRADMAFANYGPWRRMRKICVINLFSRKRAESWASVREEVNEMVQTVAGKTGSPVNIGQLVFALTRNITYRAAFGSRSHEGQGEFVKILQEFSKLFGAFNMQDFLPWLGWVHAQGFQDRMA RARKSLDVFIDKIIDDHMAKRKANTENKDDSEAADTDMVDELIAYFSDDAGKEGDDPNSGFKLTRDNIKALIMDVMFGGTETVASVIEWTMAELMKSPEDLKRVQQELTDVVGLNRRLQETDLENLTYLKCAVKESLRLHPPIPLLLHETVEDTSVAGYSFPAGSRVWINAWAIARDPTAWDEPETFKPSRFLDDGSPDFKGSNFEFIPFGSGRRSCPGMALGLYGLEMAVAHLLHCFAWELPGGMKPSELDMNDVFGLTAPKAVQLVAVPTYRLNCPL sequence no. 30 >Peach_XP_007199246.1 MDDLLLHQALESLQSSPMLFILLFLLIISWFVLFMSRRKLPYPPGPRGWPIIGNMLMMDQLTHRGLAQLAKQYGGLLHLQMGVLHIMVVSSPKVAREILQVQDSSFANRPANAAISYLTYDRADMAFANY GPFWRRMRKICVINLFSRKRAESWASVREEVEEMVRHVATKTSSPVNIGQLVFTLTKNITYRAAFGSSSHEGQGEFVKILQEFSKLFGAFNMQDFLPWLGWVHAQAFKDRMAKARRSLDVFIDKIIDDHM AKRNTNKAKKDDNEAETDMVDELIAFFSDDAAKESDDPNSTFRLTRDNIKAIIMDVMFGGTETVASVIEWTMAELMKSPEDLQKVQQELINVVGLNRRVQETDLENLTYLKCAVKESLRLHPPIPLLLHE TAEETSVAGYSFPVGSRVYINAWAIARDPTAWDEPETFKPSRFLKDGSPDFKGSDFEFLPFGSGRRSCPGMQLGLYGLEMAVAHLLHCFAWELPEGMKPNELDMNDVFGLTAPKAVQLVAVPSYRLNCPL sequence no. 31 >Peach_XP_007203643.1 MDSLQALQPLQPMTLFFIIPFLFLSGLVFLYRSRRRSPYPPGPTGLPIIGNMLMMDQLTHRGLAKLAKQYGGIFHLRMGFLHMVGISNPDVARQVLQVQDNIFSNRPATIAISYLTYDRADMAFAHYG PFWRQMRKLCVMKLFSRKRAESWDAVRDEVDTAVRTVAVQAGSAVNIGELVFSLTKNIIYRAAFGTSSEEGQDEFIGILQEFSKLFGAFNIADFIPCLGWVDPQGLNNRLARARQSLDFIDSIIDDHIQ KKKKKKSEGSNGGETDMVDELLAFYSDEAKVNESDDNLQNAINLTRDNIKAIIMDVMFGGTETVASAIEWAMAELMRNPEELKRVQQELADVVGLDRRPEEGDFEKLTYLKCALKETLRLHPPIPLLLHE TAEDAEVAGYHIPKKSRVMINSWAIGRDKDSWEDAESFKPSRFLKEGVPDFKGSNFEFLPFGSGRRSCPGMQLGLYSLELAVGHLLHCFTWELPDGMKPSELDMNDVFGLTAPRASRLIAVPSKRVVCPL sequence no. 32 >Banana_XP_009384541.1 MEWSEEVTPLHFMVCFALPLVLYVVATRRRGKLPFPPGPPQLPVIGNMLMMDQLTHRGLAKLGEHYGGLCHLRLGFLHAFAVSTPEIARQVLQVQDNVFSNRPATIAISYLTYNRADMAFAHYGPFWRQMRKLCVMKLFSKKRAESWASVREEVDVAVRSLADRAGSAVNVGELLFNLTKNIIFRAAFGTQSHENQNEFISILQEFSKLFGSFNIGDFIPWLSWMDPQGINKRLKVARA SLDRFIDKIIDEHMANRKEAADASDADMVDDMLAFLDESGYRCQAGERDDLQGTLKLTRNNIKAIIMDVMFGGTETVASAIEWAMAELMKSPEDMKRVQQELAHVVGLDRKVHESDLDKLSFLKCVTKETLRLHPPIPL LLHETAEDCEVAGYTVPARSRVMINVWAIGRDKSSWEDADAFRPSRFTPGGCAASLDFKGNYFEFLPFGSGRRSCPGMQLGLHALELAVAQLIHCFTWTLPDGMKPSELDMGDVFGLTAPRAVRLAAVPAPRLSCPLY sequence no. 33 >Banana_XP_009384087.1 MEWLEEVTSMRFVVCVVVPVTLLLAASTRWRRKLPFPPGPTPLPIVGNMLMMGQLTHRGLAKLSERFGGGLCHLRLGFVHVFAVSTSEIARQVLQVQDAVFSNRFATIAITYLTYDRADMAFAHYGPFWR QMRKLCVMKLFSKKRAVSWASVREEVDAAVRAVTDGAGAAVNLGELMFNLTKNITFRAAFGTQSHENQEEFIAILQEFSLLFGAFNIGDFIPWVSWMDLQGINKRFKVAREALDGFIDKIIEEHMANPK EADAEDSDMVDEMLAFFEESRDRTKENEADELQRTLRLTRNNIKAIIMDVMFGGTETVASAIEWAMAELMKNPEDMRRVQEELASVVGLHRKVRESDLDKLPHLKCAVKETLRLHPPIPILLHETAEDCQLTGYAVPARSRIMINVWAIGRDKSAWEDAEVFRPSRFAPGGEAAALDFKGGCFEFLPFGSGRRSCPGMQLGLHALELAVAQLTHCFSWELPDGMPKGELDMGDMFGLTPRAVRLVAVPTPRLTCPLY sequence no. 34 >Banana_XP_009411495.1 MVWVEEVTSMHLILLCLMLPLTLLLIVNIAARRRRRLPLPPGTPPLPIIGNMLLMNQLTHRGLARLAKLYGGLLHLRLGFVHHFVVSTPDVARQVLQVQDSVFSDRPTATTAIVYLTYNRSDLAFAQCG PYWRQMRKLCVTKLFSRKHAESWLSIPEEVDAAVCTVAKHAGSAFNVRDLAFLTKNIVFRSAFGKRSDENQEEYIAVVQEIATLLGAFSVGDFIPWLSWMDPQGINKRLRVARATLDFIDRIIDEHM ATDAANADMVGVMLAFLEESSHHHRQEEGDDLKGTLRLSRANIRAVMMDVMFGGTETVAIAEWALADLLTSPDDLKRVQEELAMVVGLDRKVHESHLDKLSFLKCAIKETLRLHPPFPLLLHQTADHC EVAGYSIPARSPVMINVWAIGRDESAWKDADAYRPSRFAPGGDAAALDFKGNCFEFLPFGSGRRSCPGMQLGMHELELAVAQLLHCFTWALPDGMKPTELDMGDVFGLSAPKAVPLVAVPTPRLSCPLN sequence no. 35 >Banana_XP_009403617.1 MDWFHQLSFMVASVFIPLALLSFFCMRSGRKLLLPPGPQPLPIIGNMLMMDQLTHRGLARLAERYGGLFHLRLGSLHAVVVSTPEMARLVLQVQDASFCNRPVTAAIAYLTYDRADMAFANYGPFWRQTRKLCVMKLFSRRRLQSWASVRQEVDSAVRFAARRSGSSVDVGDLAFTLAKNVTFMAAFGAQSHGNQGEFAGILQEYSKLFGEFNISDFLPWLRWMDLQGIDKRLKVARQAIDRYIDVIIDDHLANPKEADAQDADMVDGMLAFLGDSGDTNEGGDLHGDLSLTRSNIKAIIMDVMFGGTETVALGIEWAMAELLKSPEELKRTQQELASVVGLHRKVDDSDLDKLPYLKCAVKEMLRLHPPLPLLQHQATQDCELAGYFIPVGTRVFVNAWGIGRDRDAWKSPNAFRPSRFALGGDAAAFDFRGSCFELLPFGSGRRSCPGMQLGLYVLELAVAQLLHCFDWSLPAGTKPGDLDMGDVFGLTAPKAVRLMAVPTPRLTCPLL Sequence number ɔ6 >Wheat_A0A077RPS5 It should be noted that there may be some inaccuracies in the translation of "配列番号36" as "Sequence number ɔ6" due to the unclear "ɔ" character. It might be a misrepresentation in the original text. If this is a specific code or symbol with a known meaning, it should be translated more accurately according to its actual significance.MATFAKIAMELLADPLMWLFLASLALVAMQRRRLGSAPFPPGPKPLPVIGNMTLVDQLTHRGLAALAKQFGGLLHLRFGWLHVLAVSTPEYAREVLHAQDGVFSNRPATIAVVYLTYGRDMAFAHNGAYWRQMRKLCVTKIFSRRRAETWLAVREGYGALAREVGRRSGEAVNLGELIFNLTVSVIFRAAFSTCDEDGLIEFIAILQEFSRLLGLFHIGDFFPWLAWVGNRRLSTARGALDRFIDKIVDE HMRRGKDPADPDADLVDGLLGFLADANPREDALRFTRDNVKAMIMDMLFGPETVGSTTEWAMAEMMRSPDELERLQQELADVVGLDRAVEESDLDKLPFLRCVVKEALRMHPPIPVLLHEAAKDCVVGGYSIPRGSRVLVIAWAINRDCGAWKDGDTFRPARFIPGEGEAAGLDLKGSCYEFLPFGSGRRSCPAQGLGQHAVEFAVAQLAHGFNWKLPDGMKPAELDMGDIFGLTASRSTRLYAVPTPRLTCPV sequence no. 37 >Wheat_W5A2I1 MAAYAKVGTEFLKDPLIWLFLASLAFVILQRRRLGSAPFPPGPKPLPVIGNMALVDQLTHRGLAALAKQYGGLLHLRLGRLHVYAVSTPEYAREVLHVQDAALSNRPATIAVVYLTYGRSDMAFAHNG AYWRQMRKLCVTKIFSRRRAETWLAVREGYGALACAVSRRCGEAVNLGELIFNLTVSVIFRAAFSTRDEDGLDEFIAILQEFSSLLGLFHIGDFFPWLGWVGRRGFNRRLRTARGALDKFIDRIIDEHM KRGKNAADPEADLVDGLLAFLAEANPISGKHREDALRFTRDNAKAMIMDMLFGGPETVGSMTEWAMAEMMRSPDDLRRLQRELANVVGLHRTVDETDLDKLPFLRCVVKEALRMHPPIPLLLHEAAKDC FVGGYSVPKGSRVLVNAWAINRDPGAWKDGDTFRPSRFMPGEGEAAELDLNGGCYEFLPFGSGRRSCPAQGLGQHAVEFAVAQLAHGFNWELPDGMKPAELDMGDIFGLTALRATRLYAVPTPRLTCPM SEQ ID NO: 38 >Wheat_W5B6W3 MVGLAKIAMDWLQEPLSWLFVASFVFVVLQRHRQRLRGKAPPLPPGPSPLPIVGNMFMMDQLTHRGLAALARQYGGILHLRLGQVHAVVLSTPEYAQEVLQAQDVAFSNRPATVAAIYLTYDRADMAFAHYGP FWRQMRKLCVMKLFSRRRAGTWLAVRDESAALVRAVARRSGESVNLGELIFNLTKNVTFRAAFGADAAGDAGKRDEFIAIMQEFSQLFGGSIGDFIPWLGWVDQGLNVRARTARAALDEFIDKIIDEHMRRGK NPDDVDADMVDDMLAFLPEAKPKKDAGDDLQNSLHLTRDNIKAMIMDVMFGGTETVASGIEWAMTEMMHSPDDLRRLQQELADVVGLDRNVDESDLDKLPFLKCVIKETLRLHPPIPILHHENAEDCVVGGYSV PRGSSVMINVFAIGRDAKVWKDADTFRPSRFMTGEGEAARVDFKGSCFEFLPFGSGRRSCPGMALGIYSLEFAVAQLAHGSWALPDGMKPSELDMTDIFGLTAPRATRLCAVPTPRLTYPLISDVDATHKGNT SEQ ID NO: 39 >Wheat_A0A077RQ37 MVGLTKIAMEWLQDPLSWLFVASVVFVMLQRRRRRRGRAPPLPPGPNPLPIVGNMSMMDQLTHRGLTALAKKYGGFLHLRLGKVHAFAVSTPEYAQEVLQVQDAAFSNRPASLAATYLTYDRADMAFAHHGPFW RQMRKLCVMKLFSRRRPETWLAVRNESAALVRAVARRRSGETVNLGELIFNLAKNVTFRAAFGAGAAGDAGKQEEFIAILQEFSKLFVEFCIGDFIPWLSWADPQGINVRLRARAALDQFIDKIIDEHMKRGRN PDDVDADMVDDMLAFLPEARTKKAAGDRGDDLQNTLRLTRDNIKAMIMDVMFGGTETVASAIEWAMSEMMHCPDDLRRLQQELADTVGLDQNVDESDLDKLPFLKCVIKETLRLHPPIPLLNHENAEDCVVGGY SVPRGSRVMINVFAIGRDASTWKDADAFRPSRFMEGEEAAGVDFKGGGCFEFLPFGSGRRSCPGMALGLYSLELIVAQLAHGFNLALPDGMAPSELDMRDVFGLTVPRATRLCVVPTPRLTCSLVADDDAAHQA sequence no. 40 >Wheat_W5AC21 MVDLSMIDMEWLQEPLSWLFVASVIFVLLQRRRGKAPPLPPGPYSPPIVGNIFMMDQLTHRGFAALAKQYGGLLHLRLGKVHTFAVSTPEYAQQVLQAQDAAFSHRPATIATTYLTYDRADMVFARYGPFW RQMRKLCVMKLFSRRRPGTWLAVRDESAALVRAVARRSGEPVNLGDLIFNLSMNVTFRAAFGAEAAGDGDGRKQHEFIAIMQEFSKLFGAFSIGDFIPWLGWADPQGIKVRLRAARTALDEFIDKIIDEHIK RGRNPDDMDADMVDGMLAFLPEAKPDKAAGDDLHHTLLRNDNIKAIIMDVMFGGTETVASAIEWAMAEMMHSPNDLLQLQQELADTVGIDRNVDEPDLNKLSFLKCVIKETLRLHPPIPLLHRENAEDCV LGGYSVPQGSSVNINVFAMGRDTKVWKDADTFRPSRFMEGEEAAGVDFKGGGCFQFLPFGSGRRSCPGMALGLYSLELVIAQLAHGFNWALPNGEKPSVLDMSDIFGLTAPRATRLWVVPTPRLTCPLVVDV sequence no. 41 >Rice_Os03g0112900 MANGVAEYLLMDPWLLVLVLASMAFALLHLRRRARRGAPPLPGPRPLPIIGNMLMMDQLTHRGLAAMAARYGGLLHLLRLGRVHMVVVSSPEHAREVLQVQDGDFSNRPASIAIAYLTYGRADMAFSHYGHFWRQVRKLSAVRLFSRRRAQSWRAVRDESAKLVGAIARRAGEAVDLGELIFGLTKDVIFRAAFGTRDGGGHGELEVLLQEFSKLFGAFNVGDFIPWLAWLDPHGINRRRAARAALDSVIDRIIDEH VSNPAGDEDADMVDDMLAFLDEAGRDQTGGGGELQGTLRLTRDNIKAIIMDFVFGGTEVASAIEWAMAELLHSPGDLRRLQAELADVVGLGRGVEEGDLEKLPFLRCVAMETLRLHPPIPLLLHEAAADCVVGGYSVPRGARVVVNVWSVGRDAGAWKGDAGAFRPARFMAGGEAAGMDLRGGFELLPFGSGRRACPAIVLGMYELELVVARLVHAFGWAPPGGVAPEELDMADGFGLTAPRAALRAVPTPRLTCPM sequence no. 42 >Rice_Os10g0512400 MADMVKFTMEWLQDPLSLAIVVTVAVLIMRMQRRRAAPFPPGPKPLPIVGNMAMMDQLTHRGLAALAKEYGGLMHLRLGRLHAFAVSTPEYAREVLQAQDGAFSNRPATTAIAYLTYDRADMAFAHYGPFWR QMRKLCVVKLFSRRRAETWLAVRDESAALVRAVAASRGEAAVNLGELIFNLTKNVIFRAAFGTRDGEGHDEFIAILQEFSKLFGAFNIGDFIPWLSWADTNGINARLVAARTALDRFIDKIIDEHMERGKNPD DADADMVDDMLAFLAEAKPHAGKAAAAAAGAGDGADDLQNTLRLTRDNIKAIIMDVMFGGTETVASAIEWAMAEMMHSPDDLRRVQEELAAVVGLGRDVAESDLDKLPFLRCVIKETLRLHPPIPILLHETA ADCLVAGYSVPRGSRVMVNVWAIARDRAAWGPDADAFRPSRFAAGAAAEGLDFRGGCFEFLPFGSGRRSCPGMALGLYALELAVARLAHGFNWSLPDGMKPSELDMSDIFGLTAPRATRLSAVATPRLTCPLY SEQ ID NO: 43 >Maize_A0A0B4J2X1 MVTVAKIAMEWLQDPLSWVFLGTLALVVLQLRRRGKAPLPPGPKPLPIVGNMAMDQLTHRGLAALAERYGGLLHLRLGRLHAFAVSTPEYAREVLQAQDGAFSNRPATIAIAYLTYDRADMAFAHYGPFW RQMRKLCVMKLFSRRRAETWVAVRDECAALVRAVASGGGGGGEAVNLGELIFNLTKNVTFRAAFGTRDGEDQEEFIAILQEFSKLFGAFNVVDFLPWLSWMDLQGINRRLRAARSALDRFIDKIIDEHVRRG KNPDDADADMVDDMLAFFAEAKPPKKGPAAAADGDDLHNTLRLTRDNIKAIIMDVMFGTETVASAIEWAMAEMMHSPDDLRRLQQELADVVGLDRNVNESDLDKLPFLKCVIKETLRLHPPIPLLLHETA GDCVVGGYSVPRGSRVMVNVWAIGRHRASWKDADAFRPSRFTPEGEAAGLDFKGGCFEFLPFGSGRRSCPGTALGLYALELAVAQLAHGFNWSLPDGMKPSELDMGDVFGLTAPRATRLYAVPTPRLNCPLY SEQ ID NO: 44 >Maize_B4FWF9 MAAAVANIGMEWLQDPLSWVFLGTVCLVVLQQLRRRRGKAPLPPGPKPLPIVGNMGMMDQLTHRGLAALAETYGGLLHLRLGRLHAFAVSTPEYAREVLQAQDGAFSNRPATAAIAYLTYDRADMAFAHYGPF WRQMRKLCVMKLFSRRRAETWAAAVRDECAALVRAVAVGGGSGGEAVNLGELIFSLTKNVTFRAAFGTRDGEGQEEFIAILQEFSKLFGAFNVGDFLPWLGWMDLQGINRRLRARAARSALDFIDKIIDEHVRRG KSPDDADADMVDDMLAFFVEATPGKATGAAAAAADGGDDLHNTLRLTRDNIKAIIMDVMFGGTETVASAIEWAMAEMMHSPDDLRRVQQELADVVGLDRNVSESDLDRLPFLRCVIKETLRLHPPIPLLLHETA DDCVVAGYSVPRGSRVMVNVWAIGRHRASWKDADAFRPSRFAAPEGEAAGLDFKGGCFELPFGSGRRSCPGMALGLYALELAVAQLAHAFNWSLPDGMKPSEMDMGDIFGLTAPRATRLYAVPTPRNLCPLY sequence no. 45 >Soybean_G3E7M3 MANLDLDPFQTSILILVPIALLVALLSRTRRRAPYPPGPKGLPIIGNMLMMEQLTHRGLANLAKHYGGIFHLRMGFLHMVAISDPVAARQVLQVQDNIFSNRPATIAISYLTYDRADMAFAHYGPFWR QMRKLCVMKLFSRKRAESWQSVRDEVDAAVRAVASSVGKPVNIGELVFNLTKNIIYRAAFGSSSQEGQDEFIKILQEFSKLFGAFNIADFIPYLGCVDPQGLNSRLARARGALDSFIDKIIDEHVHKMK NDKSSEIVDGETDMVDELLAFYSEEAKLNNESDDLQNSIRLTKDNIKAIIMDVMFGGTETVASAIEWAMAELMRSPEDQKRVQQELADVVGLDRRAEESDFEKLTYLKCALKETLRLHPPIPLLLHETA EDATVGGYFVPRKARVMINAWAIGRDKNSWEEPETFKPARFLKPGVPDFKGSNFEFIPFGSGRRSCPGMVLGLYALELAVAHLLHCFTWELPDGMKPSEMDMGDVFGLTAPRSTRLIAVPTKRVVCPLF SEQ ID NO: 46 >Soybean_I1J8P0 MANLDLDPFQTSILILVPIALLVALLSRTRRRAPYPPGPKGLPIIGNMLMMEQLTHRGLANLAKHYGGIFHLRMGFLHMVAISDPVAARQVLQVQDNIFSNRPATIAISYLTYDRADMAFAHYGPFWR QMRKLCVMKLFSRKRAESWQSVRDEVDAAVRAVASSVGKPVNIGELVFNLTKNIIYRAAFGSSSQEGQDEFIKILQEFSKLFGAFNIADFIPYLGCVDPQGLNSRLARARGALDSFIDKIIDEHVHKMK NDKSSEIVDGETDMVDELLAFYSEEAKLNNESDDLQNSIRLTKDNIKAIIMDVMFGGTETVASAIEWAMAELMRSPEDQKRVQQELADVVGLDRRAEESDFEKLTYLKCALKETLRLHPPIPLLLHETA EDATVGGYLVPKKARVMINAWAIGRDKNSWEEPESFKPARFLKPGVPDFKGSNFEFIPFGSGRRSCPGMVLGLYALELAVAHLLHCFTWELPDGMKPSEMDMGDVFGLTAPRSTRLIAVPTKRVVCPLF SEQ ID NO: 47 >Soybean_Q2LAL3 MDWQSMMGNLDPQRTILILVPLTLLLLLLLSRTRPRPPYPPGPKGFPIIGNMFMMDQLTHRGLANLAKHYGGIFHLRMGFLHMVAISDPDAARQVLQVQDNIFSNRPATIAISYLTYDRADMAFAHYGP FWRQMRKLCVMKLFSRKRAESWQSVRDEVDAAVRAVASSVGKPVNIGELVFNLTKNIIYRAAFGSSSQEGQDEFIKILQEFSKLFGAFNIADFIPYLGCVDPQGLNSRLARARGALDSFIDKIIDEHVHK MKNDKSSEIVDGETDMVDELLAFYSEEAKLNNESDDLQNSIRLTKDNIKAIIMDVMFGGTETVASAIEWAMAELMRSPEDQKRVQQELADVVGLDRRAEESDFEKLTYLKCALKETLRLHPPIPLLLHET AEDATVGGYFVPRKARVMINAWAIGRDKNSWEEPETFKPARFLKPGVPDFKGSNFEFIPFGSGRRSCPGMVLGLYALELAVAHLLHCFTWELPDGMKPSEMDMGDVFGLTAPRSTRLIAVPTKRVVCPLF SEQ ID NO: 48 >Soybean_K7MH28 MDLLLELKTALEPFRETLLFTIPLTLLLLGIVSRIRRKTAPYPPGPKGLPLIGNMNIMNQLTHKGLANLAKQYGGVLHLRIGFLHMVAISNAEAAREVLQVQDNIFSNRPATIAISYLTYDRADMAFAH YGPFWRQMRKICVMKLFSRKRAESWNTVRDEVDFIIRSVTNNLGSPNVNVGELVFNLTKNIIYRAAFGSSSQEGQDEFISILQEFSKLFGAFNVADFVPFLGWVDPQGLNKRLVKARASLDSFIDKIIDEH VQKRRSGHDGDEESDMVDELLNFYSHEAKLNDESDELLNSISLTRDNIKAIIMDVMFGTETVASGIEWAMAELMRSPDDLRRVQQELADVVGLDRRVEESDLEKLVYLKCAVKETLRLHPPIPLLLHE TAEDAAVCGYHVPKGSRVMINAWAIGRDKSAWEDAEAFKPSRFLNPHVPDFKGSNFEFIPFGGSGRRSCPGMQLGLYTLELAMAHLLHCFTWELPDGMKPSELDTSDVFGLTAPRASRLVAVPFKRVLCPL SEQ ID NO: 49 >Soybean_I1LHY5 MDWQSMMGNLDPQRTILILVPLTLLLLLLLSRTRPRPPYPPGPKGFPIIGNMFMMDQLTHRGLANLAKHYGGIFHLRMGFLHMVAISDPDAARQVLQVQDNIFSNRPATIAISYLTYDRADMAFAHYGP FWRQMRKLCVMKLFSRKRAESWQSVRDEVDSAVRAVANSVGKPVNIGELVFNLTKNIIYRAAFGSSSQEGQDDFIKILQEFSKLFGAFNIADFIPYLGRVDPQGLNSRLARARGALDSFIDKIIDEHVQK KNNYQSSEIGDGETDMVDELLAFYGEEAKLNNESDDNLQNSIRLTKDNIKAIIMDVMFGGTETVASAIEWVMSELMRSPEDQKRVQQELADVVGLDRRVEESDFEKLTYLKCALKETLRLHPPIPLLLHE TAEDATVGGYFVPRKARVMINAWAIGRDKNSWEEPETFKPARFLKPGVPDFKGSNFEFIPFGSGRRSCPGMVLGLYALELAVAHLLHCFTWELPDGMKPSEMDMGDVFGLTAPRSTRLIAVPTKRVVCPLF SEQ ID NO:50 >Artichoke_KVI02897.1 MDVLQLQGLLHPMPILFYVALPLTFFLLSSFRRKPLPPGPRGWPLIGNMLMMDQLTHRGLASLADKYGGLLHLKMGFSHTVAVSSPEMARQVLQVQDNVFANRPATIAITYLTYDREDMAFANYGPF WRQMRKLCVMKLFSRKRAESWDSVRDEVVSMIKITAASSGSAVNLGELVFGLTHDIIYRAAFGSISHEGKEEFIRILQEYTKLFGAFNLADFIPWLGFIDPAGLNTRLPKARGELDGFIDKIIDEHLR KEKKTGDDADNDMVDEMLAFYSEEGKVNEGEDLQNAIRLTRNNIKAIIMDVMFGGTETVASAIEWALTELMHTPEALKRAQQELADVVGLDRRVEESDFEKLTYFKCIIKETLRLHPPIPVLLHQSSE ATEVAGYHIPKGTRVMVNAYAINRDKNAWKDPNTFNPSRFLENGAPDFKGSNYEFLPFGSGRRSCPGMQLGLYAMEVAHLLHSFTWQLPDGMKPSEIDMTDVFGLTAPKATRLVAVPTPRLLCPLY SEQ ID NO:51 >Artichoke_KVH92322.1 MDSIQIPISIYVIIALLTFSFLAWLRRKNLPPGPMGWPVIGNMLMMDQLTHRGLARLAEKYGGILHLKMGFRHTIAVSSPEIARQILQVQDNIFANRPATIAITYLTYDRVDMAFADYGPFWRQMR KLCVMKLFSRKRAESWDSVRDEVDSMVTTTATNSGLPVNLGELVFGLTHDIIYRAAFGSTSHEGKEEFIRILQEYTKLFGAFNLADFIPWLGFIDPAGLNTRLPAARAALDGFIDKIIDQHLAKEKK VGDENVDNDMVDEMLAFYSEEGKTNEAEDLQNAIKLTRDNIKAIIMDVMFGGTETVASAIEWAMTELMHTPEALKLVQQELTNVVGLDRRVEESDFEKLPYFKCVIKETLRLHPPIPVLLHQSSEA TEVSGYHIPKGTRVMVNSYAINRDKNAWTDPNTFNPSRFLQNGAPDFRGSNYEFLPFGSGRRSCPGMQLGLYAMEMAVVHLLHCFNWELPDGMKPSEIDMGDVFGLTAPKAIRLVAVPTPRLLCPLY SEQ ID NO:52 >Radish_GSRAST00026355001 MESSISQTLGQVLDPTTGILIVVSIFIFIGLITRRRRPPYPPGPRGWPIIGNMSMMDQLTHRGLANLAKKYGGLCHLRMGFLHMYAVSPSDVAKQVLQVQDSVFSNRPTIAISYLTYDRADMAFAHYGP FWRQMRKVCVMKVFSRKRAESWASVRDEVDKMIRSVSNNVGKSINVGEQIFALTRNITYRAAFGSACEKGQDEFIRILQEFSKLFGAFNVADFIPYFGWIDPQGINKRLVCARNDLDGFIDDIIDEHMKK KENQNTADDGEVVDTDMVDDLLAFYSEEAKLVSEATELQNSIKLTRDNIKAIIMDVMFGGTETVASAIEWALTELLRSPEDLKRVQQELAEVVGLDRHVEESDIEKLTFLKCTLKETLRLHPPIPLLLHE TAEDTEIDGYFVPKKSRVMINAFAIGRDKNSWVDPETFRPSRFLEPGVPDFKGSNFEFIPFGSGRRSCPGMQLGLYALELAVAHILHCFTWKLPDGMKPSELDMSDVFGLTAPKATRLYAVPSTRLICAV sequence no. 53 >Radish_GSRAST00007419001 MESSVSQTLSQVFDPTTTILIVVSVFIFIDLITRRRRSYPPGPRGWPIIGNMLMMDQLTHRGLANLAKKYGGLCHLRMGFLHMYAVSSPDVARQVLQVQDSIFSNRPATIAISYLTYDRADMAFAHYGP FWRQMRKVCVMKVFSRKRAESWASVRDEVDKMIRSVSSNVGKSINVGEQIFALTRNITYRAAFGSACEKGQDEFIRILQEFSKLFGAFNVADFIPYFGWIDPQGINKRLVKARNDLDGFIDDIIDEHMKK KENQNTVDDGDVDTDMVDDLLAFYSEEAKLVSETTDLQNSIKLTRDNIKAIIMDVMFGGTETVASAIEWALTELLRSPEDLKRVQQELAEVVGLDRHVEESDIEKLTFLKCTLKETLRLHPPIPLLLHE TAEDTEIDGYFVPKKSRVMINAFAIGRDPKSWPDAETFRPSRFLEPGVADFKGSNFEFIPFGGSGRRSCPGMQLGLYALELAVAHILHCFTWKLPDGMKPSELDMNDVFGLTAPRATRLFAVPSTRLICAV SEQ ID NO:54 >Radish_GSRAST00001088001 MESLLSQNLNHVIDPIPSALLITISLLVVVYLISQWLKPLYPPGPKGLPVIGNMLMVNQLTHHGLAKLANRYGGLFHLRMGFRHVFAITSPDVARQVLQVQDISFSNRPVTVAINYLTYDLADMAFAP YGPFWRQMRKVCVMKVFSRKRAESWASVREEVNNMVRSLSSNDVGKPVNVGDLIFTLTRNITYRAAFGAACETEQDEFIRILQEFSKLFGAFNIADFIPFLGWLDLQGINKRLVKARNDLDGFIDEVID EHMKKRETINGDEDTDMVDDLLAFYSEDSSPNRSKNAVKLTRDNIKALVMDVMFGGTETMASGIEWALTELLRNPAELKRLQQELTEVVGLDQRVDDTHLEKLTFLKCTLKETMRLHPPIPLILHEAIE DTKLQGFSVPKGSRLMINAFAIARDPKLWVDPETFRPCRFMEPGMPDFMGTNFEFIPFGAGRRSCPGMQLGLYAMEVAVANIIHCFTWKLPDGMKPSELDMSDVMGLTAPRATRLIAVPDTRLICSVWP SEQ ID NO: 55 >Radish_GSRAST00042054001 MDCLLCSPILYVVIFLWYLVRVLVTRGNPPPGPKGYPIIGNMKLKNQLNHRGLAELAKQYGGLLHLQMGKIHIVAASTAEMAREILQVQDVVFANRPANPAISYLTYNRADMAFANYGPLWRQMR KVCVMKLFSRKRAESWASVRDEINTMVQTLTKQTGSPVNVGELVFALTRNITYRAAFGSFARDGQDEFVKILQEFSKLFGAFDITEFPWMKWFSNRDFSKRLENARKSLDGFIDRIIDAHIEKKNS RKQEDDGLEDDMVDELMAFYSGENGGKSNDSLSTFRLTRDNIKALVMDVMFGGTETVASAIEWAMTELMKNPHELVKLQQELADVIGLNRQFHESDLENLPYFRCAMKETLRLHPPIPLLLHEAAADSVVSGYSIPRDSRVMINVYAIGRDGSVWTEPDAFRPGRFMEDKAPDFKGSDFEFLPFGSGRRSCPGMQLGLYAMELAVAHMLHSFDWKLPEGVSSGDLDMTDMFGLTAPRATRLIAVPSYRLKCPMVI sequence number 56 >Radish_rs_Aokubi_v1_EVM13601 MYTPMTLILLVPLLLFLYWHLLSRRRLRLRKSYPPGPKGLPIIGNILMMNQVNHRGLAKLSRTYGGLLHLRLGLSHLFVVSSPQIARQVLQVQDHVFSNRPTTIAIRYLTYGQSDLAFGNYGPFWRRMRKLYVMMLFSRKAESWASVDEEVHKAVRFVAANVGKPLNICKVAFSLTRDITFGAAFGSSSSTSDEGRLDEFLEIIQEFSKLFGEFNVADYVPSWLSWIDPQGINKRVERARKSLDCFIESIINDHLDKKKKTENNVDVDEETDMVDQLLAFYKEEVKVKDSETKINLNNIKGIIMDVMFGGTETVALAIEWVLTELLRSPENMKRVQEELATVVGLERWSVEDTHLEKLTFLKCVLKETLRLHPPFPLLLHETVEDAEVSGYSIPKGSRVMVNTYALGRNPNSWSDPEIFNPSRFLDPGAPDLKGNSFEFIPFGSGRRSCPGMQLGLYAFELAVAHLLHCFTWKLPNGVKPGDVDTIEGPGLTVPKANSLVAVPTTRLLSPIVLESHNV sequence number 57 >Onion_AC.SP3B.Locus_5396.1.10 MMDMQSILIFTLPFVTLLFLVITSRRRPKLPLPPGPRPLPIIGNLNLIDKLTHRGLAHLANQYGGIFHLKLGSVHTFSISTPEIAKEVLQTQDLAFSNRPATIAITYLTYDRADMAFTHYGPFWRQIRKLCVMKLFSRKRAESWASVREEIEKAVSTAAAGAGTVVNVGELVFNLTKNITFRAAFGAKSGEEQDEFLGILQEISKLFGAFNVGDFVPGLRYLDPQGIGRRMRKVRKELDGFIDRIIDEHVQNRKEVDDVEADMVDEMLAFVGQGKSIGRDSDELRLTRNNIKAIIMDVMFGGTETVASAIEWAMAELLKSPEDLKRLQQELTSVVGLDRKVQDSDLDKLPYLKCVIKETLRLHPPIPLLLHETAEDCEIQGYSIPKKSRVMINVWAIGRDKSAWKDADQFKPSRFVKGGEYEQVDFKGNFFELLPFGAGRRSCPGMQLGLYALDLTVANMAHCFDWELPDGMKPGEMDMSDVFGLTAPRAVRLAAVPSPRLTCRI DNA sequence number 58 >Lettuce_LsF5H_1_1 SEQ ID NO:59 >Lettuce_LsF5H_2_1 SEQ ID NO: 60 >Endive_ce_gene1 SEQ ID NO: 61 >Endive_ce_gene2 SEQ ID NO: 62 >Endive_ce_kethel_v0.1_EVM71979 SEQ ID NO: 63 >Chicory_ci_vitessa_fr_v1_EVM170737 SEQ ID NO: 64 >Chicory_ci_vitessa_fr_v1_EVM170780 SEQ ID NO: 65 >Chicory_ci_vitessa_fr_v1_EVM158591 SEQ ID NO: 68 >Celery_RZ_draft_99.605_EVM363634 SEQ ID NO: 69 >Celery_RZ_draft_99.605_EVM348724 SEQ ID NO: 70 >Celery_c17480_g1_i1|m.26831 SEQ ID NO:71 >Brassica_Bo1g005770 SEQ ID NO:72 >Brassica_Bo7g117840 SEQ ID NO: 73 >Brassica_Bo7g119430 SEQ ID NO:74 >Brassica_Bo3g093960 SEQ ID NO: 75 >Brassica_XP_013607401.1 SEQ ID NO:76 >Eggplant_c18725_g1_i1|m.19489 SEQ ID NO:77 >Eggplant_sm_67_3_v1_EVM112823_1_ SEQ ID NO:78 >Potato_XP_006340697.1 SEQ ID NO:79 >Apple_XP_008372753.1 SEQ ID NO: 80 >Apple_XP_008337913.1 SEQ ID NO: 81 >Apple_XP_008391587.1 SEQ ID NO:82 >Pear_XP_009378215.1 SEQ ID NO: 83 >Pear_XP_009338655.1 SEQ ID NO:84 >Pear_AGR44939.1 SEQ ID NO: 85 >Pear_XP_009346304.1 SEQ ID NO:86 >Pear_XP_009378429.1 SEQ ID NO:87 >Peach_XP_007199246.1 SEQ ID NO: 88 >Peach_XP_007203643.1 SEQ ID NO:89 >Banana_XP_009384541.1 SEQ ID NO: 90 >Banana_XP_009384087.1 SEQ ID NO: 91 >Banana_XP_009411495.1 SEQ ID NO:92 >Banana_XP_009403617.1 SEQ ID NO: 93 >Wheat_A0A077RPS5 SEQ ID NO:94 >Wheat_W5A2I1 SEQ ID NO: 95 >Wheat_W5B6W3 SEQ ID NO:96 >Wheat_A0A077RQ37 SEQ ID NO:97 >Wheat_W5AC21 SEQ ID NO: 98 >Rice_Os03g0112900 SEQ ID NO: 99 >Rice_Os10g0512400 CGAAGGATTAGGCTAACCTAAATTATTAAAGATTGAGAAGTTTAATTTACAGATATTTTTAATCCTTGAAAGAAAATCAAAGTGTGATAAATATGATTTTTTAAAAAAAACAATCACCCCTTTTCTCCAAATTATCTGCGGTAAGGCATGTCCACTTTGTGGCCACCCTCCAATGCCGAACCTTTGCTGAATCTCAAGCAAAGGAACAAGCAAGAAATCTCTGCTGGAAGGGAGGTTGATAAA AGGCACTACTCCACAAATGTCAGTTTTTTTGTCTGAGCTTGGTATTATTATTATTAAAAGTAGCTAAATTGTAAGTTGGTTACAATTGTGCAATAATAGTAATAACTTGCAACGTCCGAGCTAATTTACAATTATGGGTGACCATGTTTCGCATCACTAAGGCTGTGTTTAGTCCATACCAAAATTGAAAGTTTGGTTAAACTTGGAACGATGTGACGAAAAAGTTGAAAGTTTGTGTGTAGGAAAGTTTTG sequence number 100 >Maize_A0A0B4J2X1 SEQ ID NO: 101 >Maize_B4FWF9 SEQ ID NO: 102 >Soybean_G3E7M3 SEQ ID NO: 103 >Soybean_I1J8P0 SEQ ID NO: 104 >Soybean_Q2LAL3 SEQ ID NO: 105 >Soybean_K7MH28 SEQ ID NO: 106 >Soybean_I1LHY5 SEQ ID NO: 107 >Artichoke_KVI02897.1 SEQ ID NO: 108 >Artichoke_KVH92322.1 SEQ ID NO: 109 >Radish_GSRAST00026355001 SEQ ID NO: 110 >Radish_GSRAST00007419001 SEQ ID NO: 111 >Radish_GSRAST00001088001 SEQ ID NO: 112 >Radish_GSRAST00042054001 SEQ ID NO: 113 >Radish_rs_Aokubi_v1_EVM13601 SEQ ID NO: 114 >Onion_AC.SP3B.Locus_5396.1.10 SEQ ID NO: 115 > Lettuce_LsF5H_1_1 CDS WT SEQ ID NO: 116 >Lettuce_LsF5H_2_1 CDS WT SEQ ID NO: 174 > Lettuce_LsF5H_1_1 CDS mutation 1 SEQ ID NO: 175 >Lettuce_LsF5H_2_1 CDS mutation 2 SEQ ID NO: 176 >Lettuce_LsF5H_2_1 CDS mutation 3 SEQ ID NO: 177 >Lettuce_LsF5H_2_1 CDS mutation 4 SEQ ID NO: 178 >Lettuce_LsF5H_2_1 CDS mutation 5 SEQ ID NO: 179 >Lettuce_LsF5H_2_1 CDS mutation 6

Claims

1. 1. A lettuce plant (Lactuca sativa) comprising a modified F5H1 gene and a modified F5H2 gene, the genes comprising a modification compared to their corresponding wild-type F5H genes, the wild-type F5H1 gene having SEQ ID NO: 115 and the wild-type F5H2 gene having SEQ ID NO: 116, wherein the modification in the F5H1 gene is a C→T mutation at position 370 of SEQ ID NO: 115, and the modification in the F5H1 gene and the modification in the F5H2 gene result in reduced or absent protein expression and / or activity of an F5H1 protein and an F5H2 protein compared to the expression of proteins produced by the corresponding wild-type F5H1 gene and wild-type F5H2 gene, and the presence of the modified F5H1 gene and the modified F5H2 genes in the lettuce plant results in reduced wound-induced surface discolouration compared to a lettuce plant not comprising the modified F5H1 gene and the modified F5H2 gene.

2. 2. The lettuce plant of claim 1, wherein the modified F5H1 gene is homozygous and the modified F5H2 gene is either heterozygous or homozygous.

3. 3. The lettuce plant of claim 1, wherein the modified F5H2 gene encodes a protein having one or more amino acid substitutions.

4. 4. A part of a lettuce plant according to any one of claims 1 to 3, wherein the part is a leaf, a whole head of the plant, a fruit, an inflorescence, a seed, a curd, a stem, a tuber, a bulb or a root, or a processed form thereof.

5. 5. A food product comprising a part of a lettuce plant according to claim 4.

6. A seed capable of developing a lettuce plant according to any one of claims 1 to 3.

7. A seed of the lettuce plant according to any one of claims 1 to 3, comprising a modified F5H1 gene and a modified F5H2 gene in its genome.

8. 4. A seed from which the lettuce plant of any one of claims 1 to 3 can be grown and / or derived from said lettuce plant, wherein the seed is selected from the group consisting of microspores, pollen, ovaries, ovules, embryos, embryo sacs, egg cells, cuttings, roots, root caps, hypocotyls, cotyledons, stems, leaves, flowers, anthers, seeds, meristematic cells, protoplasts and cells, or tissue cultures thereof.

9. 1. A method of producing a lettuce plant that exhibits reduced wound-induced surface discolouration, comprising reducing the endogenous amount of F5H1 protein in the lettuce plant, wherein the endogenous amount of F5H1 protein in a lettuce plant is reduced by mutating a wild-type F5H1 gene to produce a modified F5H1 gene, wherein the modified F5H1 gene comprises a mutation C to T at position 370 of SEQ ID NO:

115. method.

10. 10. The method of claim 9, wherein the mutagenesis is performed by CRISPR, chemical reagents, radiation, or a combination thereof.

11. 11. A lettuce plant comprising reduced F5H1 expression, wherein the reduction is achieved by the method of claim 9 or 10.

12. 1. A method for selecting lettuce plants that exhibit reduced wound-induced surface discolouration, the method comprising screening a lettuce plant or a population of lettuce plants for the presence of the modified F5H1 gene of claim 1, wherein the modified F5H1 gene comprises a modification that results in a premature stop codon and, optionally, applying a phenotypic test to identify lettuce plants that exhibit reduced wound-induced surface discolouration, and selecting lettuce plants that exhibit reduced wound-induced surface discolouration.

13. 1. A method of producing a lettuce plant that exhibits reduced wound-induced surface discolouration, comprising: (a) crossing a lettuce plant containing the modified F5H1 gene and the modified F5H2 gene of claim 1 with another lettuce plant; (b) optionally, performing one or more self-crosses and / or out-crosses; and (c) optionally selecting lettuce plants containing the reduced wound-induced surface discoloration after each round of selfing or outcrossing. A method comprising:

14. 14. The method of claim 13, wherein the lettuce plants are selected phenotypically and / or by the use of molecular markers.

15. 10. A method for producing seeds of a hybrid lettuce plant, comprising crossing a first parent lettuce plant with a second parent lettuce plant and harvesting seeds of the resulting lettuce plant, wherein the first parent lettuce plant and / or the second parent lettuce plant comprise the modified F5H1 gene and the modified F5H2 gene of claim 1.

16. 10. A method for determining the presence of the modified F5H1 gene and the modified F5H2 gene of claim 1 in a lettuce plant, the method comprising the steps of obtaining a nucleic acid sample from the lettuce plant, comparing the nucleic acid with a nucleic acid sample obtained from a control lettuce plant comprising a wild-type F5H1 gene and a wild-type F5H2 gene, and detecting a polymorphism between the two nucleic acid samples, wherein the detected polymorphism is indicative of the presence of the modified F5H1 gene and the modified F5H2 gene.

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