Melon plants resistant to ToLCNDV-ES

Abstract

The present invention relates to muskmelon plants that are ToLCNDV-ES resistant to a resistant ToLCNDV-ES strain, and methods of producing such plants. The TIR-NBS-LRR gene is identified as conferring resistance.

Description

[0001] This application relates to a melon plant (Cucumis melo) resistant to infection with the Spanish strain of tomato leaf curl New Delhi virus (ToLCNDV-ES). In one aspect, the resistant melon plant possesses a genomic introgression segment on chromosome 5 that confers resistance to ToLCNDV-ES in a dominant manner. Markers for identifying those segments and methods for identifying or producing resistant melon plants are also disclosed. Furthermore, potential causal genes for QTLs are provided, along with methods for selecting or modifying causal genes. In one aspect, the resistant melon plant contains a modified endogenous gene conferring resistance.

[0002] Tomato leaf curl virus (ToLCNDV) is a two-component bean golden mosaic virus that is transmitted by the whitefly, specifically the tobacco whitefly (Bemisiatabaci Genn.) (Hemiptera: Amycidae), and causes severe symptoms such as yellowing and curling of leaves and stunting of plants in susceptible hosts.

[0003] Prior to 2016, ToLCNDV was confined to India and other Asian countries, where it was predominantly found on Solanaceae hosts, particularly tomatoes. However, it has since been reported in Spain and other Mediterranean basin countries, where it is mostly found on Cucurbitaceae hosts and appears to have adapted to them (Fortes et al., Viruses 2016, 8, 307; doi:10.3390 / v8110307). DNA-A sequence analysis revealed that the Spanish isolate (ToLCNDV-ES) belongs to a different phylogenetic cluster than other ToLCNDV isolates.

[0004] The Mediterranean ToLCNDV isolate (also known as the European ToLCNDV isolate) is called ToLCNDV-ES, and all ToLCNDV-ES isolates are now known to be genetically highly identical (>99% identity, Juarez et al., 2019, Natural hosts and genetic diversity of the emerging tomato leaf curl NewDelhi virus in Spain. Frontiers Microbiology, 10, 140. https: / / doi.org / 10.3389 / fmicb.2019.00140), and are distinct from all other ToLCNDV isolates found outside of Europe. Although ToLCNDV-ES is infectious to tomatoes (Ruiz et al., 2017, Biological characterization of Tomato leaf curl New Delhi virus from Spain. Plant Pathology, 66, 376–382), ToLCNDV-ES appears to be poorly adapted to this host; instead, it infects Cucurbitaceae plants such as squash, melons, cucumbers, and pumpkins very effectively.

[0005] This genetic consistency among European isolates is also thought to explain their preference for and adaptation to Cucurbitaceae hosts.

[0006] WO2018 / 011075 describes a QTL on chromosome 5 of the melon genome that confers resistance to ToLCNDV. The cultivar Coliseo F1 (Nunhems), containing the QTL, has also been commercialized. Wild donors used for introgression of the QTL had small fruits (not exceeding 6 cm in length) with white, tart flesh. The donors had a mean ToLCNDV disease score of 7.4 on a scale of 1 = plant death to 9 = asymptomatic, while the mean disease score of the QTL-backcrossed improved lines was approximately 5.1. The QTL was mapped to the region defined by SNP_01 to SNP_06. Seeds were deposited with NCIMB 42585, representing BC4S4 cultivars containing homozygous introgression fragments, where the donor nucleotides were homozygous for SNP_01 to SNP_06. Later, the TolCNDV resistance was finely located between SNP_03 and SNP_04, and two SNP markers were added between these two markers, named SNP_07 and SNP_08, see US11591611B2 which is incorporated herein by reference.

[0007] The resistance conferred by the aforementioned QTL was found to be disrupted by ToLCNDV-ES isolates in the Spanish field.

[0008] One object of the present invention is to provide a QTL on chromosome 5 to confer resistance to the destructive ToLCNDV-ES strain.

[0009] This invention discloses melon plant cells and melon plants resistant to infection by the current TolCNDV-ES strain present in Spain.

[0010] It is well known that ToLCNDV-ES can indeed infect a wide variety of plant species within the Cucurbitaceae family, including melon species. It is also known that ToLCNDV-ES is continuously spread from infected to uninfected plants by the plant-sucking pest, the tobacco whitefly (whitefly). Transfer of ToLCNDV-ES from one crop species to another, or even from weed species, has been demonstrated. Whiteflies can acquire ToLCNDV-ES from outside the control area, even from different species, and transfer it to melon plants growing within the control area. Therefore, whitefly-borne control has limited effectiveness in preventing ToLCNDV-ES infection. ToLCNDV-ES resistant melon plants offer the advantage that even if plants surrounding the melon-growing area are infected with ToLCNDV-ES, they will withstand ToLCNDV-ES infection without significant yield loss.

[0011] The inventors have discovered a novel, wild-type ToLCNDV-ES resistance donor germplasm for the hairy melon subspecies (C. melo ssp. melo), which remains asymptomatic (average score 9.0) against the most aggressive ToLCNDV-ES strain in Spain, which causes more severe disease symptoms on previously resistant melon plants containing QTLs as described in WO2018 / 011075 and US11591611B2.

[0012] When resistance from the selected donor line (where resistance in the original germplasm is highly variable) was mapped in the F2 population derived from a cross between a new donor and a susceptible melon breeding line, it was surprising to find that the QTL was mapped to the same region on chromosome 5 as described in WO2018 / 011075 and US11591611B2. However, when tested with the latest ToLCNDV-ES strain, this region was significantly different from previous QTLs on chromosome 5 due to differences in SNP haplotypes (see [link to ToLCNDV-ES strain]). Figure 3And because of their high average disease scores, QTLs from new donors are able to withstand infection by the latest ToLCNDV-ES strains. Therefore, the latest strains are also referred to as "resistance-destroying strains" because they can overcome the resistance of WO2018 / 011075. This does not mean that the resistance of the QTLs described in WO2018 / 011075 is completely lost due to the new strains, but the resistance becomes less effective because the disease symptoms caused by the new ToLCNDV-ES strains are more severe, especially under high viral stress.

[0013] To distinguish QTL5 from WO2018 / 011075 (and US11591611B2), this article names QTL5 QTL5rb, which stands for "resistance-destroying QTL5," meaning that QTL5rb confers resistance against the most aggressive ToLCNDV-ES strain.

[0014] QTL5rb confers resistance in a single-gene dominant manner.

[0015] The chromosome 5 region located by QTL5rb contains the largest NBS-LRR gene cluster in melon (González et al., BMC Genomics 2014, 15:1131, title: Interspecific and intraspecific genevariability in a 1-Mb region containing the highest density of NBS-LRR genes found in the melon genome), but this paper was able to identify causal genes by using various elimination methods.

[0016] The potential causal gene was located and found to encode a TIR-NBS-LRR resistance protein. This resistance protein was compared with the TIR-NBS-LRR protein in a previous QTL5 (WO2018 / 011075) and found to be different. When compared by a pairwise comparison (using Needle), the two proteins shared 95.2% sequence identity with each other. Furthermore, when compared with the corresponding protein in the reference genome (melonomics.net, DHL92 version 4) and the susceptible variety Vedantrais, the protein of the present invention shared 98.6% sequence identity with the protein in the reference genome and the protein in Vedantrais. The protein is identical in the reference genome and the Vedantrais variety.

[0017] The quantitative trait locus (QTL) (QTL5rb) on chromosome 5 was introgressed from wild-type donor germplasm into improved cultivated melon lines. In accordance with the Budapest Treaty, Nunhems BV has deposited seeds of selected wild-type donors with fixed QTL5rb introgression and homozygous donor SNP nucleotides under accession number NCIMB 44139. Among these seeds, donor genotypes possessing the single nucleotide polymorphism (SNP) markers (SNP_01 to SNP_07) described herein were present.

[0018] Wild donors are inconsistent and not agronomically valuable germplasm. By identifying QTL5rb and transferring it from the donor to cultivated melons, it is now possible to produce high-agronomical-value cultivated melon varieties and cultivars resistant to the destructive ToLCNDV-ES strain (with consistent characteristics and marketable fruit with high Brix content and good shelf life), and thus these melon varieties can be cultivated in ToLCNDV-ES-infested areas without yield loss.

[0019] "Melon plant cell" or "melon plant" is also referred to in the art as melon plant cell or melon plant, and in the context of this invention, it should be understood as a plant cell derived from the species melon or a plant belonging to the species melon.

[0020] Melons can be classified into three types: Roman melon (C. melo cantalupensis), smooth-skinned melon (C. meloinodorous), and netted melon (C. melo reticulatus). Roman melons, also known as cantaloupes, are primarily round with prominent ribs and little netting. Most have sweet, orange flesh and are typically very fragrant. North American 'cantaloupes' are not this type, but rather belong to the true muskmelon category, unlike European cantaloupes. Smooth-skinned melons (or winter melons) can be further subdivided into different types, such as honeydew melon, Spanish melon, sugar melon, and Japanese melon. Netted melons are true muskmelons with a netted rind (netted structure) and include Gallia melon, Salle melon, and North American cantaloupe.

[0021] Melon and its wild relatives are diploid and have 12 pairs of homologous chromosomes, numbered 1 to 12.

[0022] In the context of this invention, "resistance" or "resistance to..." should be understood to mean a plant that is a host species of a particular pathogen and can therefore be infected by a given pathogen, but wherein the plant contains a genetic element (e.g., an introgression fragment) that, compared to a susceptible plant lacking that genetic element, results in reduced pathogen growth and / or spread in the infected plant. In the context of this invention, "resistance" or "resistance to..." specifically refers to plant cells or the plant being resistant to ToLCNDV-ES, particularly to the most aggressive ToLCKEV-ES strains, such as those that destroy resistance. Resistance is a relative term and can span a range of (different) responses triggered by pathogen infection in plant cells or the plant. The effects of those responses in plant cells or the plant can be measured by various means. Typically, the effect is measured by defining the level of symptoms appearing in plant cells or the plant. Typically, the average symptoms of several plants (e.g., 10 or more) of a strain are compared to the average symptoms of several plants of a control strain or variety, preferably a susceptible control strain or variety. Therefore, at least 10 or more individual plants of a strain or variety are scored at a point in time, and an average disease score is calculated.

[0023] Regarding this invention, based on phenotypic observations obtained after ToLCNDV-ES infection, the following known symptom levels are applied:

[0024] 1 = Plant death

[0025] 2 = Severe mosaic virus, leaf curling, chlorosis, and stunted growth. No recovery.

[0026] 3 = Severe mosaic virus and leaf curling, yellowing, and stunted growth. No recovery.

[0027] 4 = Leaf curling and mosaic virus, chlorosis, no growth decline or slight growth reduction. No recovery.

[0028] 5 = Leaf curling and mosaic virus, chlorosis, no growth decline. The upper part of the plant has slightly recovered.

[0029] 6 = Mild curling, mosaic virus, and chlorosis; no growth decline. The upper and middle regions of the plant have recovered.

[0030] 7 = Mild curling, mosaic virus, and chlorosis; no stunted growth. Symptoms only appear in the lower part of the plant.

[0031] 8 = Slight mosaic disease

[0032] 9 = Asymptomatic

[0033] To determine the symptom level (or disease score), it is preferable to infect young plants with ToLCNDV-ES. Young plants are preferably seedlings at the stage where the first true leaf has unfolded, preferably plants approximately 12-15 days after sowing. Infection is preferably carried out by feeding a virus-carrying vector (whitefly). For this purpose, the plants germinate and grow under optimal or near-optimal conditions. The symptom level is preferably determined at least once, for example, 30 or 40 days after infection (or later, such as 31, 32, 33, 34, 35, 40, or 41 days after infection). Optionally, the symptom level is determined two or even three times at different time points after infection to confirm the results, for example, a first score at approximately 15, 20, or 25 days after ToLCNDV-ES infection and a second score at approximately 30 or 40 days (or later) after infection. See also the examples. In one respect, when grown under the same conditions and infected in the same manner, a plant line is said to be resistant to ToLCNDV-ES infection if it has a mean disease score of at least 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, or higher, such as at least 9.0, while a susceptible control line or control variety, such as the variety Gandalf (or Gandalf F1, Nunhems BV) or Vedantrais, has a mean disease score of 2.5 or lower, or 2.0 or lower. In one respect, a plant line containing a homozygous or heterozygous QTL5rb is said to be resistant to ToLCNDV-ES infection if it has a higher average disease score to a destructive ToLCNDV-ES strain than a plant containing the QTL5 of US11,591,611, such as NCIMB42585 or the cultivar Coliseo F1 (Nunhems).

[0034] It has been observed that QTL5rb, located on chromosome 5 from a wild melon donor plant, infiltrates into cultivated melon plants, conferring resistance to ToLCNDV-ES infection in the cultivated plants or cells derived from them. The presence of the corresponding fragment in a heterozygous state alone is sufficient to confer ToLCNDV-ES resistance, demonstrating that this fragment confers resistance in a dominant manner. Single nucleotide polymorphisms (SNPs) on chromosome 5 tightly linked to the ToLCNDV-ES-conferring QTL5rb were identified. Furthermore, a potential causal gene for QTL5rb has been identified. The SNP nucleotides of the resistance donor (i.e., the nucleotides of the infiltrated fragment) are present in homozygous form in preserved seeds; specifically, for SNP_01 to SNP_07, the donor nucleotides are present in homozygous form (linked to QTL5rb). Therefore, SNPs can be used to test for the presence of introgressive fragments containing QTL5rb in plant cells, plant tissues, plant parts, and / or marker-assisted selection (MAS) to transfer QTLs into improved melon lines or varieties. SNPs can also be used to select plants containing introgressive fragments smaller than those present in preserved seeds, whereby the smaller subfractions retain QTL5rb. For example, it was found that backcross lines with resistant donor nucleotides for SNP_03, SNP_04, and SNP_05 (but not SNP_01, SNP_02, SNP_06, and SNP_07) all retained QTL5rb, see Examples. Alternatively, one or more potential causal gene sequences of SNPs and / or QTL5rb can be used to identify other donors containing QTL5rb and to introgress QTLs into cultivated melons or to transfer QTL5rb from one cultivated melon plant to another.

[0035] Therefore, in one aspect, the present invention relates to cultivated melon plant cells or melon plants containing an introgression fragment from a ToLCNDV-ES resistant donor plant on chromosome 5, wherein the introgression fragment comprises a sequence of the donor plant between SNP_01 and SNP_07, preferably a sequence of the donor plant between SNP_01 and SNP_06, or between SNP_01 and SNP_05, or between SNP_01 and SNP_04, or between SNP_02 and SNP_06, or between SNP_03 and SNP_05, more preferably a sequence of the donor plant between SNP_01 and SNP_04, or between SNP_02 and SNP_04, or between SNP_03 and SNP_04. The QTL5rb conferring ToLCNDV-ES resistance is present on the introgression fragment, as can be determined by resistance assays as described herein.

[0036] Therefore, in one aspect, the present invention relates to melon plant cells or melon plants containing an introgression fragment on chromosome 5 derived from (obtained from, available from) a ToLCNDV-ES resistance donor plant, wherein the introgression fragment contains a sequence of a ToLCNDV-ES resistance donor melon plant, the sequence being selected from the following markers:

[0037] -SNP_01 and SNP_07, and comprising the following donor SNP nucleotides: SNP_02 (i.e., cytosine at nucleotide 242 of SEQ ID NO: 2 or guanine in the complementary strand), SNP_03 (i.e., guanine at nucleotide 81 of SEQ ID NO: 3), SNP_04 (i.e., adenine at nucleotide 101 of SEQ ID NO: 4 or thymine in the complementary strand), SNP_05 (thymine at nucleotide 101 of SEQ ID NO: 5 or adenine in the complementary strand) and / or SNP_06 (adenine at nucleotide 101 of SEQ ID NO: 6 or thymine in the complementary strand); optionally, donor nucleotides of SNP_01 and / or SNP_07 may be present, i.e., cytosine at nucleotide 81 of SEQ ID NO: 1 (SNP_01) and / or cytosine at nucleotide 81 of SEQ ID NO: 7 (SNP_07).

[0038] -SNP_01 and SNP_06 and contain the following donor SNP nucleotides: SNP_02 (i.e., cytosine at nucleotide 242 of SEQ ID NO: 2 or guanine in the complementary strand), SNP_03 (i.e., guanine at nucleotide 81 of SEQ ID NO: 3), SNP_04 (i.e., adenine at nucleotide 101 of SEQ ID NO: 4 or thymine in the complementary strand) and / or SNP_05 (thymine at nucleotide 101 of SEQ ID NO: 5 or adenine in the complementary strand); optionally, donor nucleotides of SNP_01 and / or SNP_06 may be present, i.e., cytosine at nucleotide 81 of SEQ ID NO: 1 (SNP_01) and / or adenine at nucleotide 101 of SEQ ID NO: 6 or thymine in the complementary strand (SNP_06).

[0039] -SNP_01 and SNP_05 and comprising the following donor SNP nucleotides: SNP_02 (i.e., cytosine at nucleotide 242 of SEQ ID NO: 2 or guanine in the complementary strand), SNP_03 (i.e., guanine at nucleotide 81 of SEQ ID NO: 3) and / or SNP_04 (i.e., adenine at nucleotide 101 of SEQ ID NO: 4 or thymine in the complementary strand); optionally, donor nucleotides of SNP_01 and / or SNP_05 may be present, i.e., cytosine at nucleotide 81 of SEQ ID NO: 1 (SNP_01) and / or thymine at nucleotide 101 of SEQ ID NO: 5 or adenine in the complementary strand (SNP_05).

[0040] -SNP_02 and SNP_07, and comprising the following donor SNP nucleotides: SNP_03 (i.e., guanine at nucleotide 81 of SEQ ID NO: 3), SNP_04 (i.e., adenine at nucleotide 101 of SEQ ID NO: 4 or thymine in the complementary strand) and / or SNP_05 (thymine at nucleotide 101 of SEQ ID NO: 5 or adenine in the complementary strand) and / or SNP_06 (adenine at nucleotide 101 of SEQ ID NO: 6 or thymine in the complementary strand); optionally, donor nucleotides of SNP_02 and / or SNP_07 may be present, i.e., cytosine at nucleotide 242 of SEQ ID NO: 2 or guanine in the complementary strand (SNP_02) and / or cytosine at nucleotide 81 of SEQ ID NO: 7 (SNP_07).

[0041] -SNP_02 and SNP_06 and contain the following donor SNP nucleotides: SNP_03 (i.e., guanine at nucleotide 81 of SEQ ID NO: 3), SNP_04 (i.e., adenine at nucleotide 101 of SEQ ID NO: 4 or thymine in the complementary strand), and / or SNP_05 (thymine at nucleotide 101 of SEQ ID NO: 5 or adenine in the complementary strand); optionally, donor nucleotides of SNP_02 and / or SNP_06 may be present, i.e., cytosine at nucleotide 242 of SEQ ID NO: 2 or guanine in the complementary strand (SNP_02) and / or adenine at nucleotide 101 of SEQ ID NO: 6 or thymine in the complementary strand (SNP_06).

[0042] -SNP_02 and SNP_05 and contain the following donor SNP nucleotides: SNP_03 (i.e., guanine at nucleotide 81 of SEQ ID NO: 3) and / or SNP_04 (i.e., adenine at nucleotide 101 of SEQ ID NO: 4 or thymine in the complementary strand); optionally, donor nucleotides of SNP_02 and / or SNP_05 may be present, i.e., cytosine at nucleotide 242 of SEQ ID NO: 2 or guanine in the complementary strand (SNP_02) and / or thymine at nucleotide 101 of SEQ ID NO: 5 or adenine in the complementary strand (SNP_05).

[0043] -SNP_03 and SNP_07, and comprising the following donor SNP nucleotides: SNP_04 (i.e., adenine at nucleotide 101 of SEQ ID NO: 4 or thymine in the complementary strand) and / or SNP_05 (thymine at nucleotide 101 of SEQ ID NO: 5 or adenine in the complementary strand) and / or SNP_06 (adenine at nucleotide 101 of SEQ ID NO: 6 or thymine in the complementary strand); optionally, donor nucleotides of SNP_03 and / or SNP_07 may be present, i.e., guanine at nucleotide 81 of SEQ ID NO: 3 (SNP_03) and / or cytosine at nucleotide 81 of SEQ ID NO: 7 (SNP_07).

[0044] -SNP_03 and SNP_06 and contain the following donor SNP nucleotides: SNP_04 (i.e., adenine at nucleotide 101 of SEQ ID NO: 4 or thymine in the complementary strand) and / or SNP_05 (thymine at nucleotide 101 of SEQ ID NO: 5 or adenine in the complementary strand); optionally, donor nucleotides of SNP_03 and / or SNP_06 may be present, i.e., guanine at nucleotide 81 of SEQ ID NO: 3 (SNP_03) and / or adenine at nucleotide 101 of SEQ ID NO: 6 or thymine in the complementary strand (SNP_06).

[0045] -SNP_03 and SNP_05 and contain the following donor SNP nucleotides: SNP_04 (i.e., adenine at nucleotide 101 of SEQ ID NO: 4 or thymine in the complementary strand); optionally, donor nucleotides of SNP_03 and / or SNP_05 may be present, i.e., guanine at nucleotide 81 of SEQ ID NO: 3 (SNP_03) and / or thymine at nucleotide 101 of SEQ ID NO: 5 or adenine in the complementary strand (SNP_05).

[0046] -SNP_03 and SNP_04 and the donor SNP nucleotide (i.e., guanine) of SNP_03 contained at nucleotide 81 of SEQ ID NO: 3, and / or the donor SNP nucleotide containing SNP_04, i.e., adenine at nucleotide 101 of SEQ ID NO: 4 or thymine in the complementary strand.

[0047] The term "between" does not exclude the first and / or last mentioned SNP nucleotide as a donor nucleotide; therefore, for example, "between SNP_01 and SNP_07" in one respect refers to SNP_01 and / or SNP_07 containing the resistance donor nucleotide. Resistance donor nucleotides are shown, for example, in Table 1 of this document. However, the first and last mentioned SNPs can also be different nucleotides, such as nucleotides of a recurrent parent. Furthermore, when referring to SEQ ID NO:, in one respect also covers variations of the mentioned sequence, i.e., the mentioned SNP nucleotide at the equivalent nucleotide position in a sequence having at least 95%, 96%, 97%, 98%, or 99% sequence identity with the mentioned SEQ ID NO. Therefore, slight variations may exist in the nucleotides before or after the mentioned SNP nucleotide.

[0048] Therefore, when this document refers to an introgression fragment containing a sequence “between two SNPs (single nucleotide polymorphisms)”, this in one aspect encompasses either or both of the two SNPs originating from a resistance donor, i.e., having a donor nucleotide at the SNP location. In another aspect, both SNPs originate from a receptor, such as in a susceptible melon plant, while only the region between the two SNPs originates from a resistance donor and confers ToLCNDV-ES resistance, i.e., a resistance-conferring donor fragment containing QTL5rb is located between the two SNP markers. Thus, for example, a plant may contain an introgression fragment of a melon plant containing a ToLCNDV-ES resistance donor sequence between SNP_03 and SNP_04, in one aspect of which such a plant contains guanine (G) at nucleotide 81 of SEQ ID NO: 3 and / or adenine (A) (or thymine in the complementary strand) at nucleotide 101 of SEQ ID NO: 4, i.e., donor nucleotides. On the other hand, only the region between these two SNPs (the entire region or a part thereof) comes from the donor, while SNP_03 and SNP_04 come from the recipient.

[0049] Therefore, regarding QTL5rb on chromosome 5, SNP_01, SNP_02, SNP_03, SNP_04, SNP_05, SNP_06, and SNP_07 can all possess a resistance donor haplotype (or genotype). Alternatively, only SNP_03 and SNP_04 can possess a resistance donor haplotype (or genotype). Or only a single SNP, i.e., only SNP_03, or only SNP_04, possesses a resistance donor nucleotide. Therefore, SNPs without a resistance donor nucleotide possess another nucleotide, such as a recipient nucleotide. The recipient nucleotide of an SNP can be any of the other three nucleotides; for example, for SNP_03, the recipient nucleotide can be adenine, cytosine, or thymine. Therefore, for example, when stating that the introgression fragment is between SNP_03 and SNP_04 regarding QTL5rb on chromosome 5, both SNP_03 and SNP_04 can possess a resistance donor haplotype (or genotype). Alternatively, only a single SNP, i.e., only SNP_03 or only SNP_04, may have a resistance donor nucleotide; or even neither SNP_03 nor SNP_04 may have a resistance donor nucleotide, but the sequence between them still contains QTL5rb.

[0050] In one aspect, QTL5rb contains a gene encoding a TIR-NBS-LRR protein (“resistance protein” or resistance-conferring protein) (a potential causal gene of QTL5rb), said protein having at least 96%, 97%, 98%, or 99% sequence identity with the protein of SEQ ID NO: 8, and said protein preferably containing one or all of the following four amino acid changes compared to the susceptibility protein of SEQ ID NO: 10 (or the susceptibility protein having at least 98%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.7%, 98.8%, 98.9%, or 99%, or 99.5% or 99.8% sequence identity with SEQ ID NO: 10):

[0051] 1. At the beginning of the protein, two S (serine) residues are inserted after amino acid E9 (glutamic acid number 9); that is, the protein contains 9 serine residues (S) after E9, and amino acids 10 to 18 are serine residues in proteins having at least 96%, 97%, 98%, or 99% sequence identity with the protein of SEQ ID NO: 8 (while SEQ ID NO: 10 contains only 7 serine residues, i.e., amino acids 10 to 16); see also Figure 2 Change number 1;

[0052] 2. At the equivalent position in a protein having at least 96%, 97%, 98%, or 99% sequence identity with the protein of SEQ ID NO: 8, amino acid V483 (valine 483 of SEQ ID NO: 10) is changed to M (methionine); in SEQ ID NO: 8, the equivalent position is M484; see also Figure 2 Change number 2;

[0053] 3. In proteins having at least 96%, 97%, 98%, or 99% sequence identity with the protein of SEQ ID NO: 8, amino acid E628 (glutamic acid 628 of SEQ ID NO: 10) is changed to K (lysine); in SEQ ID NO: 8, the equivalent position is K629; see also Figure 2 Change number 3;

[0054] 4. At the equivalent position in a protein having at least 96%, 97%, 98%, or 99% sequence identity with the protein of SEQ ID NO: 8, amino acid Y678 (tyrosine 678 of SEQ ID NO: 10) is changed to H (histidine); in SEQ ID NO: 8, the equivalent position is H679, see [link to relevant documentation]. Figure 2 Change 4; or at the equivalent position in a protein having at least 96%, 97%, 98%, or 99% sequence identity with the protein of SEQ ID NO: 8, amino acid Y678 (tyrosine 678 of SEQ ID NO: 10) is changed to D (aspartic acid); in SEQ ID NO: 8, the equivalent position is D679, see [link to SEQ ID NO: 8]. Figure 1 or Figure 2 Change number 4.

[0055] In other words, QTL5rb contains a gene encoding the TIR-NBS-LRR protein (“resistance protein” or resistance-conferring protein) of SEQ ID NO: 8 or a protein having at least 96%, 97%, 98%, 99%, or 100% sequence identity with the protein of SEQ ID NO: 8 (QTL5rb potential causal gene), and said protein preferably contains one or more or all of the following amino acids:

[0056] - Nine serines (S) at equivalent positions in amino acids 10 to 18 of SEQ ID NO: 8, or in a sequence having at least 96% sequence identity with SEQ ID NO: 8.

[0057] - Methionine (M) at amino acid number 484 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0058] - Lysine (K) at amino acid number 629 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0059] - Histidine (H) or aspartic acid (D) at amino acid number 679 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8.

[0060] Two TIR-NBS-LRR proteins, one from QTL5rb and the other from QTL5 (whose resistance has been disrupted by the new ToLCNDV-ES strain), share 95.2% sequence identity with each other. However, they exhibit the aforementioned four common variations compared to proteins in the reference genome of melon and to proteins in the susceptible variety Vendretais (these two "susceptible proteins" are 100% identical). Therefore, one or all of these four common variations must be the cause of the transformation of the susceptible protein into a resistant protein. The susceptible protein can be genetically edited or mutagenized to introduce each of the four variations or a combination thereof (or all four variations) to verify the effect of each variation and its combination on ToLCNDV-ES resistance. Alternatively, proteins encoding SEQ ID NO: 8 or proteins having at least 96%, 97%, 98%, or 99% sequence identity with SEQ ID NO: 8 can be screened from wild-type donors, preferably genes containing one or all of the four variations. The effects of potential gene-edited or mutated genes in QTL5rb, or other wild-type donors containing such genes, on ToLCNDV-ES resistance can be analyzed in disease assays.

[0061] The reason why not all SNPs linked to QTL5rb as presented in this paper need to have a resistance donor genotype is that introgressive fragments containing QTL5rb from a donor may be smaller than the chromosomal fragment located to the QTL5rb region in this paper, but that fragment still contains QTL5rb. Plants containing introgressive fragments (containing QTL5rb) can still be identified by phenotype, and / or by transferring the fragment to a susceptible plant and thereby transferring the ToLCNDV-ES resistance phenotype, or by sequencing the region between SNP markers, or by other methods known to those skilled in the art, such as saturating the region with more SNP markers, allelic testing, etc. The presence and / or nucleotide sequence of causal genes between SNP_03 and SNP_04 can also be examined.

[0062] Therefore, a combination of methods can be used to confirm the presence of QTL5rb in plant cells or plants, even if the donor SNP genotype is not present for all linked SNPs. QTL5rb, when transferred to susceptible lines or varieties, confers at least a mean ToLCNDV-ES resistance of 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, or 9.0 and is dominant.

[0063] In a preferred embodiment of the invention, an infiltrated fragment of chromosome 5 from the donor plant, comprising sequences of the donor plant between SNP_01 and SNP_07, between SNP_01 and SNP_06, between SNP_01 and SNP_05, between SNP_01 and SNP_04, between SNP_02 and SNP_07, between SNP_02 and SNP_06, between SNP_02 and SNP_05, between SNP_02 and SNP_04, between SNP_03 and SNP_07, between SNP_03 and SNP_06, between SNP_03 and SNP_05, preferably between SNP_03 and SNP_04, confers resistance to ToLCNDV-ES to cultivated melon plant cells or cultivated melon plants according to the invention.

[0064] In one aspect, the introgression fragment (and / or QTL5rb and / or the potential causal gene of QTL5rb) may be obtained from (or obtained from, or may be derived from, or may be derived from) a seed or its progeny deposited under accession number NCIMB 44139, said progeny retaining QTL5rb as present in the deposited seed.

[0065] In one aspect, the introgression fragment contains donor nucleotides of SNP_03 and SNP_04, as well as QTL5rb. In another aspect, the introgression fragment contains donor nucleotides of SNP_03, SNP_04, and SNP_05, as well as QTL5rb. In another aspect, the fragment may be obtained from NCIMB 44139, its progeny (offspring) or its parent generation, or from another wild-type donor as described herein.

[0066] Therefore, in one respect, QTL5rb can be obtained from seeds whose representative samples have been deposited with accession number NCIMB 44139 or from the progeny of its retained QTL5rb, or from another wild donor (e.g., another wild melon donor) containing the following SNP marker haplotypes:

[0067] - Guanine at nucleotide 81 of SEQ ID NO: 3, or at an equivalent nucleotide in a sequence having at least 95% sequence identity with SEQ ID NO: 3;

[0068] - Adenine at nucleotide 101 of SEQ ID NO: 4, or at an equivalent nucleotide in a sequence having at least 95% sequence identity with SEQ ID NO: 4;

[0069] - Thymine at nucleotide 101 of SEQ ID NO: 5, or at an equivalent nucleotide in a sequence having at least 95% sequence identity with SEQ ID NO: 5.

[0070] Since the resistance is precisely localized between SNP_03 and SNP_04, and the causal gene encoding the TIR-NBS-LRR protein of SEQ ID NO: 8 is located between SNP_03 and SNP_04, one aspect of this document is that the infiltrated fragment contains a donor sequence between SNP_03 and SNP_04 and / or contains a gene encoding the protein of SEQ ID NO: 8 or a protein encoding a protein having at least 96%, 97%, 98%, or 99% sequence identity with the protein of SEQ ID NO: 8, and said protein preferably contains one or all of the following four amino acid changes compared to the protein of SEQ ID NO: 10:

[0071] - At the beginning of the protein, two S (serine) residues are inserted after amino acid E9 (glutamic acid number 9); that is, the protein contains 9 serine residues (S) after E9, and amino acids 10 to 18 are serine residues in proteins that have at least 96%, 97%, 98%, or 99% sequence identity with the protein of SEQ ID NO: 8 (while SEQ ID NO: 10 contains only 7 serine residues, i.e., amino acids 10 to 16); see also Figure 2 Change number 1;

[0072] - At the equivalent position in proteins having at least 96%, 97%, 98%, or 99% sequence identity with the protein of SEQ ID NO: 8, amino acid V483 (valine 483 of SEQ ID NO: 10) is changed to M (methionine); in SEQ ID NO: 8, the equivalent position is M484; see also Figure 2 Change number 2;

[0073] - At the equivalent position in proteins having at least 96%, 97%, 98%, or 99% sequence identity with the protein of SEQ ID NO: 8, amino acid E628 (glutamic acid 628 of SEQ ID NO: 10) is changed to K (lysine); in SEQ ID NO: 8, the equivalent position is K629; see also Figure 2 Change number 3;

[0074] - At the equivalent position in proteins having at least 96%, 97%, 98%, or 99% sequence identity with the protein of SEQ ID NO: 8, amino acid Y678 (tyrosine 678 of SEQ ID NO: 10) is changed to H (histidine); in SEQ ID NO: 8, the equivalent position is H679, see [link to relevant documentation]. Figure 2 Change 4; or at the equivalent position in a protein having at least 96%, 97%, 98%, or 99% sequence identity with the protein of SEQ ID NO: 8, amino acid Y678 (tyrosine 678 of SEQ ID NO: 10) is changed to D (aspartic acid); in SEQ ID NO: 8, the equivalent position is D679, see [link to SEQ ID NO: 8]. Figure 1 or Figure 2 Change number 4.

[0075] In other words, in one aspect, the infiltrated fragment comprises a gene encoding the TIR-NBS-LRR protein (“resistance protein” or “resistance-conferring protein”) of SEQ ID NO: 8 or a protein having at least 96%, 97%, 98%, 99%, or 100% sequence identity with the protein of SEQ ID NO: 8, and said protein preferably comprises one or more or all of the following amino acids:

[0076] - Nine serines (S) at equivalent positions in amino acids 10 to 18 of SEQ ID NO: 8, or in a sequence having at least 96% sequence identity with SEQ ID NO: 8.

[0077] - Methionine (M) at amino acid number 484 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0078] - Lysine (K) at amino acid number 629 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0079] - Histidine (H) or aspartic acid (D) at amino acid number 679 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8.

[0080] Preferably, the melon plant cells according to the invention are derived from cultivated melon plants, or the melon plant according to the invention is a cultivated melon plant, not a wild melon plant. Preferably, the donor is a wild melon plant. In another aspect, as described elsewhere herein, the endogenous gene encoding a "susceptibility protein" is modified to encode the resistance protein of SEQ ID NO: 8 or a protein having at least 96%, 97%, 98%, 99%, or 100% sequence identity with the protein of SEQ ID NO: 8, and said protein preferably comprises, for example, those described above. Figure 1 and Figure 2 One or more of the four variations mentioned in the text.

[0081] In one aspect, the invention relates to cultivated melon plant cells or melon plants (or plant parts, such as cells or tissues) containing an introgression fragment from a ToLCNDV-ES resistant donor plant on chromosome 5, wherein the introgression fragment confers ToLCNDV-ES resistance (i.e., it contains QTL5rb and / or the potential causal gene QTL5rb) and the introgression fragment can be detected by SNP haplotype analysis against one or more of the following SNPs from the donor plant (including the SNP haplotype): S NP_01, SNP_02, SNP_03, SNP_04, SNP_05, SNP_06 and / or SNP_07, and optionally any SNP between SNP_01 and SNP_07; in one aspect, the infiltrated fragment can be detected by SNP haplotype detection (including the SNP haplotype) against one or more (or all) of the following SNPs from the donor plant: SNP_02, SNP_03, SNP_04, SNP_05 and / or SNP_06, and optionally... Any SNP between SNP_02 and SNP_06; in another aspect, the infiltrated fragment can be detected by SNP haplotypes of donor plants for one or more (or all) of the following SNPs (including the SNP haplotype): SNP_02, SNP_03, SNP_04 and / or SNP_05, and optionally any SNP between SNP_02 and SNP_05; in another aspect, the infiltrated fragment can be detected by donors for one or more (or all) of the following SNPs. Plant SNP haplotype detection (including the SNP haplotype): SNP_03, SNP_04 and / or SNP_05, and optionally any SNP between SNP_03 and SNP_05; in another different aspect, the infiltrated fragment can be detected by SNP haplotype detection (including the SNP haplotype) of a donor plant for one or more of the following SNPs: SNP_03 and / or SNP_04, and optionally any SNP between SNP_03 and SNP_04.

[0082] Therefore, in one aspect, the plant, plant part, or plant cell contains a QTL5rb (and / or a potential causal gene of QTL5rb) and contains at least SNP donor haplotypes of SNP_03 and / or SNP_04, because these SNPs are located flanking the causal gene (QTL5rb) conferring resistance to ToLCNDV-ES. In another aspect, the plant, plant part, or plant cell contains at least SNP_03 and SNP_04, or at least SNP_03 and SNP_02, or at least SNP_04 and SNP_05 SNP donor haplotypes. Optionally, the plant, plant part, or plant cell contains a QTL5rb and contains at least SNP_02, SNP_03, and SNP_04; or at least SNP_03, SNP_04, and SNP_05 SNP donor haplotypes.

[0083] Therefore, the infiltrated fragment may contain donor SNP haplotypes of all SNP markers linked to QTL5rb (as in the seeds preserved herein), or a smaller fragment in which one or more SNP markers are absent. As further described below, even all or all but one of the donor SNP markers may be absent, while QTL5rb remains present on the infiltrated fragment.

[0084] The nucleotide sequences SEQ ID NO: 1 to SEQ ID NO: 7 containing the SNPs (referred to as SNP_01 to SNP_07) provided herein are nucleotide sequences of resistance donors, i.e., they contain donor SNP nucleotides. Therefore, in one aspect, the present invention relates to cultivated melon plant cells or melon plants (or plant parts) containing an introgression fragment from a ToLCNDV-ES resistance donor plant on chromosome 5, wherein the introgression fragment confers ToLCNDV-ES resistance and the introgression fragment can be detected by the following (including the following):

[0085] -Cytosine at nucleotide 81 of SEQ ID NO: 1 or at an equivalent nucleotide of a sequence having basic sequence identity with SEQ ID NO: 1.

[0086] And / or

[0087] - SEQ ID NO: 2 or cytosine at nucleotide 242 of SEQ ID NO: 2 (or guanine at nucleotide 242 of the complementary strand of SEQ ID NO: 2) or cytosine at an equivalent nucleotide of a sequence having basic sequence identity with SEQ ID NO: 2 (or guanine in the complementary strand).

[0088] And / or

[0089] -Guine at nucleotide 81 of SEQ ID NO: 3 or guanine at an equivalent nucleotide of a sequence having basic sequence identity with SEQ ID NO: 3.

[0090] And / or

[0091] - Adenine at nucleotide 101 of SEQ ID NO: 4 (or thymine at nucleotide 101 in the complementary strand of SEQ ID NO: 4) or adenine at an equivalent nucleotide of a sequence having basic sequence identity with SEQ ID NO: 4.

[0092] And / or

[0093] - Thymine at nucleotide 101 of SEQ ID NO: 5 (or adenine at nucleotide 101 in the complementary strand of SEQ ID NO: 5) or thymine at an equivalent nucleotide of a sequence having basic sequence identity with SEQ ID NO: 5.

[0094] And / or

[0095] - Adenine at nucleotide 101 of SEQ ID NO: 6 (or thymine at nucleotide 101 in the complementary strand of SEQ ID NO: 6) or adenine at an equivalent nucleotide of a sequence having basic sequence identity with SEQ ID NO: 6.

[0096] And / or

[0097] - SEQ ID NO: 7 or cytosine at nucleotide 101 of SEQ ID NO: 7 or cytosine at an equivalent nucleotide of a sequence having basic sequence identity with SEQ ID NO: 7.

[0098] The term "at an equivalent nucleotide (or position) in a sequence having basic sequence identity with a given sequence" refers to a SNP nucleotide in a variant sequence that, when compared as a control pair (using, for example, the Needles program with default parameters), has at least 95%, 96%, 97%, 98%, or 99% sequence identity with a given sequence. Similarly, "at an equivalent position in a sequence having basic sequence identity with a given sequence" refers to an amino acid in a variant sequence that, when compared as a control pair, has at least 95%, 96%, 97%, 98%, or 99% sequence identity with a given sequence.

[0099] In one aspect, a (cultivated) melon plant or a portion thereof is provided, the (cultivated) melon plant or the portion thereof comprising a recombinant chromosome 5, wherein the recombinant chromosome 5 comprising the introgression fragment comprises a quantitative trait locus (QTL) named QTL5rb between a cytosine of SNP_01 at nucleotide 81 of SEQ ID NO: 1 (or a cytosine at an equivalent position in a sequence having at least 95% identity with SEQ ID NO: 1) and a cytosine of SNP_07 at nucleotide 81 of SEQ ID NO: 7 (or a cytosine at an equivalent position in a sequence having at least 95% identity with SEQ ID NO: 7), said QTL5rb conferring resistance to New Delhi Tomato Leaf Curl Virus (ToLCNDV-ES) to the melon plant when present in homozygous or heterozygous form, and wherein said introgression fragment comprises one or more single nucleotide polymorphism (SNP) markers from the following group:

[0100] - Cytosine at nucleotide 242 of SEQ ID NO: 2 or at an equivalent nucleotide in a sequence having at least 95%, 96%, 97%, 98% or 99% sequence identity with SEQ ID NO: 2 (or the complementary strand) at SNP_02 (or guanine at nucleotide 242 of the complementary strand of SEQ ID NO: 2).

[0101] - Guanine at nucleotide 81 of SEQ ID NO: 3 or at an equivalent nucleotide in a sequence having at least 95%, 96%, 97%, 98%, or 99% sequence identity with SEQ ID NO: 3,

[0102] - Adenine at nucleotide 101 of SEQ ID NO: 4 or thymine at nucleotide 101 of the equivalent nucleotide in a sequence having at least 95%, 96%, 97%, 98% or 99% sequence identity with SEQ ID NO: 4 (or its complementary sequence).

[0103] - Thymine at nucleotide 101 of SEQ ID NO: 5 or at an equivalent nucleotide in a sequence having at least 95%, 96%, 97%, 98% or 99% sequence identity with SEQ ID NO: 5 (or complementary sequence) (or adenine at nucleotide 101 of the complementary strand of SEQ ID NO: 5).

[0104] - Adenine at nucleotide 101 of SEQ ID NO: 6 or thymine at nucleotide 101 of the complementary strand of SEQ ID NO: 6 or at an equivalent nucleotide in a sequence having at least 95%, 96%, 97%, 98% or 99% sequence identity with SEQ ID NO: 6 (or its complementary sequence).

[0105] Therefore, in another aspect, a (cultivated) melon plant or a portion thereof is provided, the (cultivated) melon plant or the portion thereof comprising a recombinant chromosome 5, wherein the recombinant chromosome 5 comprising the introgression fragment comprises a quantitative trait locus (QTL) named QTL5rb between a cytosine of SNP_01 at nucleotide 81 of SEQ ID NO: 1 (or a cytosine at an equivalent position in a sequence having at least 95% identity with SEQ ID NO: 1) and a cytosine of SNP_07 at nucleotide 81 of SEQ ID NO: 7 (or a cytosine at an equivalent position in a sequence having at least 95% identity with SEQ ID NO: 7), said QTL5rb conferring resistance to New Delhi Tomato Leaf Curl Virus (ToLCNDV-ES) to the melon plant when present in homozygous or heterozygous form, and wherein said introgression fragment comprises one or more single nucleotide polymorphism (SNP) markers from the following group:

[0106] - Guanine at nucleotide 81 of SEQ ID NO: 3 or at an equivalent nucleotide in a sequence having at least 95%, 96%, 97%, 98%, or 99% sequence identity with SEQ ID NO: 3,

[0107] - Adenine at nucleotide 101 of SEQ ID NO: 4 or thymine at nucleotide 101 of the equivalent nucleotide in a sequence having at least 95%, 96%, 97%, 98% or 99% sequence identity with SEQ ID NO: 4 (or the complementary strand).

[0108] - Thymine at nucleotide 101 of SEQ ID NO: 5 or at an equivalent nucleotide in a sequence having at least 95%, 96%, 97%, 98% or 99% sequence identity with SEQ ID NO: 5 (or the complementary strand) at SNP_05 (or adenine at nucleotide 101 of the complementary strand of SEQ ID NO: 5).

[0109] In one aspect, a (cultivated) melon plant or a portion thereof comprising a recombinant chromosome 5 is provided, wherein the recombinant chromosome 5 comprising the introgression fragment contains a quantitative trait locus (QTL) named QTL5rb, which confers resistance to New Delhi Tomato Leaf Curl Virus (ToLCNDV-ES) to the melon plant when present in homozygous or heterozygous form, and wherein the introgression fragment comprises one or more single nucleotide polymorphism (SNP) markers from the following group:

[0110] - Guanine at nucleotide 81 of SEQ ID NO: 3 or at an equivalent nucleotide in a sequence having at least 95%, 96%, 97%, 98%, or 99% sequence identity with SEQ ID NO: 3,

[0111] - Adenine at nucleotide 101 of SEQ ID NO: 4 or thymine at nucleotide 101 of the equivalent nucleotide in a sequence having at least 95%, 96%, 97%, 98% or 99% sequence identity with SEQ ID NO: 4 (or the complementary strand).

[0112] - Thymine at nucleotide 101 of SEQ ID NO: 5 or at an equivalent nucleotide in a sequence having at least 95%, 96%, 97%, 98% or 99% sequence identity with SEQ ID NO: 5 (or the complementary strand) at SNP_05 (or adenine at nucleotide 101 of the complementary strand of SEQ ID NO: 5).

[0113] In the aforementioned plants, QTL5rb is located, for example, in the region between the guanine of SNP_03 at nucleotide 81 of SEQ ID NO: 3 (or the guanine at an equivalent position in a sequence having at least 95% identity with SEQ ID NO: 3) and the thymine of SNP_05 at nucleotide 101 of SEQ ID NO: 5 (or the adenine at nucleotide 101 in the complementary strand of SEQ ID NO: 5 or the adenine at an equivalent position in a sequence having at least 95% sequence identity with the complementary strand of SEQ ID NO: 5).

[0114] In one aspect, in the aforementioned plants, QTL5rb is located, for example, in the region between the guanine of SNP_03 at nucleotide 81 of SEQ ID NO: 3 (or the guanine at an equivalent position in a sequence having at least 95% sequence identity with SEQ ID NO: 3) and the adenine of SNP_04 at nucleotide 101 of SEQ ID NO: 4, or the adenine at an equivalent position in a sequence having at least 95% sequence identity with SEQ ID NO: 4 (or the thymine at nucleotide 101 in the complementary strand of SEQ ID NO: 4, or the thymine at an equivalent position in a sequence having at least 95% sequence identity with the complementary strand of SEQ ID NO: 4).

[0115] In one aspect of this specification, for any SNP, the SNP nucleotide mentioned herein may also (alternatively or alternatively) be referred to by the nucleotide position on chromosome 5 of the reference genome, as shown, for example, in Table 2.

[0116] The QTL5rb that confers resistance to ToLCNDV-ES is present on the infiltrated fragment.

[0117] When the sequences provided in this paper for Seq ID NO: 2 (containing SNP_02), SEQ ID NO: 4 (containing SNP_04), SEQ ID NO: 5 (containing SNP_05), and SEQ ID NO: 6 (containing SNP_06) are compared with the sequence orientation given for the reference genome of melon, these sequences are the (-) strand / reverse strand sequences of double-stranded DNA on chromosome 5.

[0118] Therefore, another way to refer to a sequence and SNP nucleotide is to refer to the (+) strand / forward strand, which is the complementary nucleotide or complementary sequence of the (-) strand. Thus, for example, SNP_02 is cytosine at nucleotide 242 of SEQ ID NO: 2 or guanine at nucleotide 242 of the complementary strand of SEQ ID NO: 2. Therefore, in one aspect, SNP_02 is cytosine at nucleotide 242 of SEQ ID NO: 2 (or guanine at nucleotide 242 of the complementary strand of SEQ ID NO: 2) or cytosine at an equivalent nucleotide of a sequence having basic sequence identity with SEQ ID NO: 2 (or guanine in the complementary strand). Similarly, SNP_04 is adenine at nucleotide 101 of SEQ ID NO: 4 (or thymine at nucleotide 101 of the complementary strand of SEQ ID NO: 4) or adenine at an equivalent nucleotide of a sequence having basic sequence identity with SEQ ID NO: 4; SNP_05 is thymine at nucleotide 101 of SEQ ID NO: 5 (or adenine at nucleotide 101 of the complementary strand of SEQ ID NO: 5) or thymine at an equivalent nucleotide of a sequence having basic sequence identity with SEQ ID NO: 5; and SNP_06 is adenine at nucleotide 101 of SEQ ID NO: 6 (or thymine at nucleotide 101 of the complementary strand of SEQ ID NO: 6) or adenine at an equivalent nucleotide of a sequence having basic sequence identity with SEQ ID NO: 6.

[0119] In connection with this invention, "donor plant cell" or "donor plant" should refer to melon plant cells or melon plants resistant to ToLCNDV-ES, particularly to strains that disrupt resistance. Similarly, the term DNA fragment or introgression fragment from a donor plant or donor cell should refer to a fragment of chromosome 5 of a ToLCNDV-ES-resistant melon plant, whereby this fragment, when transferred to a ToLCNDV-ES-susceptible melon plant, confers ToLCNDV-ES resistance. In a preferred embodiment of the invention, the donor plant is a wild species or wild germplasm of melon. In a particular embodiment, the DNA fragment or introgression fragment from the donor plant cell or donor plant is a donor fragment obtained (or obtainable from, derived from, or present in) a plant grown from seeds preserved in NCIMB 44139, or a descendant of a plant grown from seeds preserved in NCIMB 44139, or obtained by hybridization with a plant grown from seeds preserved in NCIMB 44139. Similarly, in one aspect, the QTL5rb or QTL5rb potential causal gene from donor plant cells or donor plants is obtained (or may be obtained, or derived from, or may be derived from, or exists in) the QTL5rb or QTL5rb potential causal gene of a plant grown from seeds preserved in NCIMB 44139, or obtained from the offspring of a plant grown from seeds preserved in NCIMB 44139, or obtained by hybridization with a plant grown from seeds preserved in NCIMB 44139.

[0120] Donor melon plants can be obtained from a variety of other sources. Those skilled in the art know how to detect ToLCNDV-ES resistant donor plants containing QTL5rb, especially other sources of donor plants resistant to novel ToLCNDV-ES strains that destroy resistance. To detect such sources of ToLCNDV-ES resistant donor plants, essentially, melon plants or germplasm (e.g., wild melon plants or germplasm) can be infected with ToLCNDV-ES (especially strains that destroy resistance) by mechanical means as described by Lopez et al. (2015, Euphytica 204(3), 679-691) or by infection with the virus via whiteflies. Preferably, in the context of this invention, infection is carried out via whitefly infection. Plants that show reduced symptom levels compared to susceptible controls can then be selected and used as sources of genomic fragments or sequences conferring ToLCNDV-ES resistance. Preferred methods for infecting melon plants with ToLCNDV-ES and methods for determining symptom levels in infected plants are given under “General Methods”.

[0121] In the context of this invention, the donor plant preferably has an average symptom level of 7.7 or higher, even more preferably 8.0 or 8.5 or higher, and most preferably 9.0 or higher.

[0122] In one aspect, the donor plant contains one or more of the donor SNP haplotypes or genotypes of SNP_01, SNP_2, SNP_3, SNP_04, SNP_05, SNP_06, and SNP_07 as shown in Tables 1 and 2. In another aspect, the donor plant contains one or more of SNP_03, SNP_04, and / or SNP_05 as shown in Tables 1 and 2, or donor SNP haplotypes or genotypes of SNP_03 and SNP_04. Preferably, the SNP donor genotype is homozygous. The donor herein is, for example, a wild melon that has no agronomical value, such as producing acidic fruit.

[0123] As mentioned above, potential donor melon plants (e.g., wild germplasm from a seed bank) can also be screened for the presence of one or more SNP markers indicating the presence of QTL5rb, and / or for the presence of TIR-NBS-LRR resistance proteins, i.e., proteins encoding SEQ ID NO: 8 or encoding proteins having at least 96% sequence identity with SEQ ID NO: 8 and preferably containing proteins present in the resistance protein but not in the susceptibility protein, as described in this article. Figure 2 The presence of genes for proteins showing one or more of the four variations described above is used to screen for them.

[0124] The terms "recurrent plant cells," "recurrent plants," or "recipient plants" in connection with this invention should be understood as susceptible to ToLCNDV-ES infection, particularly to ToLCNDV-ES strains that disrupt resistance (used herein synonymously with susceptible), or resistant to ToLCNDV-ES infection, or resistant to ToLCNDV-ES infection significantly less than that of resistant donor melon plant cells or melon plants. Whether a plant is susceptible to ToLCNDV-ES infection (particularly to strains that disrupt resistance), or resistant to it, or resistant to it significantly less, can be determined by observing the symptom level following ToLCNDV-ES infection (particularly to strains that disrupt resistance). For example, recurrent plants have an average symptom level below 3.0, or equal to or below 2.5, or equal to or below 2.0. Symptom levels and methods for infecting melon plants with ToLCNDV-ES are described elsewhere herein and are accordingly applicable here. In a preferred embodiment of the invention, the recurrent melon plant cells according to the invention are derived from cultivated melon plants, or the recurrent melon plants according to the invention are cultivated melon plants. Preferably, it is a superior strain, a bred strain, or a variety. In one aspect, a parental strain containing QTL5 can also be used as a recurrent parent, and QTL5rb is introduced at a locus on chromosome 5, essentially replacing QTL5 with QTL5rb.

[0125] The terms "modified plant cells," "modified plants," "mutated or mutant plant cells," or "mutated or mutant plants" in connection with this invention should be understood as melon plant cells or melon plants in which the endogenous TIR-NBS-LRR protein on chromosome 5 has been modified or mutated, thereby making the plant less susceptible to or resistant to ToLCNDV-ES infection, particularly to resistance-disrupting ToLCNDV-ES strains, for example, exhibiting significantly higher resistance to ToLCNDV-ES infection compared to unmodified or unmutated plants. Whether a plant is less susceptible to ToLCNDV-ES infection (especially resistance-disrupting strains) or whether it is resistant or has significantly higher resistance can be determined by observing the symptom levels of modified or mutant plants after ToLCNDV-ES infection (especially infection with resistance-disrupting strains) compared to unmodified or unmutated plants. Symptom levels and methods for infecting melon plants with ToLCNDV-ES are described elsewhere herein and are accordingly applicable here. In a preferred embodiment of the invention, the modified or mutated melon plant cell or plant according to the invention is a cultivated melon plant, wherein the endogenous TIR-NBS-LRR gene encoding a susceptible TIR-NBS-LRR protein (e.g., the protein of SEQ ID NO: 10, or a susceptible protein having at least 98%, 98.5%, or 99% sequence identity with SEQ ID NO: 10) on chromosome 5 has been modified or mutated to encode a resistant TIR-NBS-LRR protein (e.g., the protein of SEQ ID NO: 8, or a protein having at least 96%, 97%, 98%, or 99% sequence identity with SEQ ID NO: 8) and preferably includes the protein described herein and, for example... Figure 2 (Proteins that have undergone one or more of the four variations shown). Preferably, it is a superior strain, a bred strain, or a variety. Thus, the plant cell or the plant contains, as defined elsewhere herein, a “modified TIR-NBS-LRR gene” in its genome.

[0126] "Introgression" refers to a segment, portion, or region of chromosome from another plant of the same or closely related species that has been introduced through hybridization or conventional breeding techniques. Introgression of a fragment from a donor plant into a recurrent plant introduces a phenotype not present in the recurrent plant into the offspring of a hybridization between the donor and recurrent plants. In this invention, the phenotype transferred from the donor plant to the recurrent plant is resistance to ToLCNDV-ES (especially resistance-destroying strains), for example, with an average disease score of at least 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, or 9.0. To introgress a fragment into a specific breeding line or variety, one or more backcrosses can be performed after the first hybridization step, for example, with the intended breeding line or variety. As understood herein, introgression can refer to the first cross between a ToLCNDV-ES resistant donor plant and a recurrent plant susceptible to or with low resistance to ToLCNDV-ES (e.g., with the presence of QTL5), and the subsequent backcrossing one or more times between the ToLCNDV-ES resistant plant obtained from the first cross and the recipient plant into which ToLCNDV-ES resistance should be introgressed. In such cases, the introgressed fragment is the result of a breeding method referred to in the verb “introgress” (such as backcrossing) into the recipient variety or cultivar. Therefore, introgression of ToLCNDV-ES resistance into recurrent plants is a human-guided technical process. Specifically, introgression as used herein refers to an artificial breeding process or method. One or more of the molecular markers (SNP markers) and / or QTL5rb and / or the potential causal genes of QTL5rb provided herein can be used in this method, for example, to help select plants containing QTL5rb. The resulting plants, i.e., cultivars or varieties containing an introgressed fragment from the donor (on chromosome 5), i.e., cultivars containing recombinant chromosome 5, are also artificial and do not exist in nature.

[0127] The infiltrated fragment can be large, such as even half a chromosome, but is preferably small, such as about 15 Mb or less, such as about 10 Mb or less, about 9 Mb or less, about 8 Mb or less, about 7 Mb or less, about 6 Mb or less, about 5 Mb or less, about 4 Mb or less, about 3 Mb or less, about 2 Mb or less, about 1 Mb (equal to 1,000,000 base pairs or less) or about 0.8 Mb (equal to 800,000 base pairs) or less.

[0128] Introgression fragments can originate from wild melon plants, wild melon germplasm, or wild relatives or local varieties of melons (donors). Wild melon plants, wild melon germplasm, or wild relatives or local varieties of melon plants can be used to introgress fragments of the donor genome into the genome of cultivated melon (Cucumis melo) to produce breeding lines or varieties with favorable agronomical characteristics. Therefore, such cultivated melon plants possess a "cultivated melon genome" but contain donor fragments, such as fragments from the genome of related wild melon genuses, such as the short-haired melon subspecies (Cucumis melo ssp. agrestis), the long-haired melon subspecies (C. melo ssp. melo), the sour melon subspecies (C. melo ssp. acidulous), the Minon melon (C. callosus), the trigonus melon (C. trigonus), the picrocarpus melon, or another wild melon or a wild relative of melon. It should be understood that the term "infiltrated fragment" never includes an entire chromosome, but only a portion of it. Therefore, the chromosome carrying the infiltration also contains one or more portions of the recurrent (recipient) melon plant, and additionally portions of the donor melon plant.

[0129] When the cultivar's chromosome 5 contains an introgression fragment, this therefore means that the cultivar plant contains a recombinant chromosome 5, and the introgression fragment contains the QTL5rb that confers resistance to ToLCNDV-ES. As described elsewhere, the introgression fragment from the donor may contain one or more or all of the donor SNP nucleotides (for SNP_01, SNP_02, SNP_03, SNP_04, SNP_05, SNP_06 and / or SNP_07) or contain one or more or all of the sequence of donor SNP nucleotides (SEQ ID NO: 1, 2, 3, 4, 5, 6 and / or SEQ ID NO: 7).

[0130] In one aspect, the introgression fragment contains a causal gene conferring resistance to QTL5rb (and can be detected by the presence of the causal gene), i.e., the introgression fragment contains a gene encoding the TIR-NBS-LRR resistance protein of SEQ ID NO: 8 or a protein having at least 96%, 97%, 98%, or 99% sequence identity with the protein of SEQ ID NO: 8, and said protein preferably contains one or all of the following four amino acid changes compared to the protein of SEQ ID NO: 10:

[0131] - At the beginning of the protein, two S (serine) residues are inserted after amino acid E9 (glutamic acid number 9); that is, the protein contains 9 serine residues (S) after E9, and amino acids 10 to 18 are serine residues in proteins that have at least 96%, 97%, 98%, or 99% sequence identity with the protein of SEQ ID NO: 8 (while SEQ ID NO: 10 contains only 7 serine residues, i.e., amino acids 10 to 16); see also Figure 2 Change number 1;

[0132] - At the equivalent position in proteins having at least 96%, 97%, 98%, or 99% sequence identity with the protein of SEQ ID NO: 8, amino acid V483 (valine 483 of SEQ ID NO: 10) is changed to M (methionine); in SEQ ID NO: 8, the equivalent position is M484; see also Figure 2 Change number 2;

[0133] - At the equivalent position in proteins having at least 96%, 97%, 98%, or 99% sequence identity with the protein of SEQ ID NO: 8, amino acid E628 (glutamic acid 628 of SEQ ID NO: 10) is changed to K (lysine); in SEQ ID NO: 8, the equivalent position is K629; see also Figure 2 Change number 3;

[0134] - At the equivalent position in proteins having at least 96%, 97%, 98%, or 99% sequence identity with the protein of SEQ ID NO: 8, amino acid Y678 (tyrosine 678 of SEQ ID NO: 10) is changed to H (histidine); in SEQ ID NO: 8, the equivalent position is H679, see [link to relevant documentation]. Figure 2 Change 4; or at the equivalent position in a protein having at least 96%, 97%, 98%, or 99% sequence identity with the protein of SEQ ID NO: 8, amino acid Y678 (tyrosine 678 of SEQ ID NO: 10) is changed to D (aspartic acid); in SEQ ID NO: 8, the equivalent position is D679, see [link to SEQ ID NO: 8]. Figure 1 or Figure 2 Change number 4.

[0135] In other words, the introgression fragment contains a causal gene conferring resistance to QTL5rb (and can be detected by the presence of the causal gene), i.e., the introgression fragment contains a gene encoding the TIR-NBS-LRR resistance protein of SEQ ID NO: 8 or a protein having at least 96%, 97%, 98%, or 99% sequence identity with the protein of SEQ ID NO: 8, and said protein preferably contains one or more or all of the following amino acids:

[0136] - Nine serines (S) at equivalent positions in amino acids 10 to 18 of SEQ ID NO: 8, or in a sequence having at least 96% sequence identity with SEQ ID NO: 8.

[0137] - Methionine (M) at amino acid number 484 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0138] - Lysine (K) at amino acid number 629 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0139] - Histidine (H) or aspartic acid (D) at amino acid number 679 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8.

[0140] Therefore, for example, in one aspect, the introgression fragment (homozygous or heterozygous form) may contain one or more or all of the following SNP genotypes or haplotypes, and can be detected by one or more or all of the following SNP genotypes or haplotypes:

[0141] - The CC genotype (homozygous for the donor nucleotide) or CX haplotype (heterozygous for the donor nucleotide) of SNP_01 at nucleotide 81 of SEQ ID NO: 1 (or an equivalent nucleotide in a sequence having at least 95%, 96%, 97%, 98% or 99% sequence identity with SEQ ID NO: 1).

[0142] - The CC genotype or CX haplotype of SNP_02 at nucleotide 242 of SEQ ID NO: 2 (or an equivalent nucleotide in a sequence having at least 95%, 96%, 97%, 98%, or 99% sequence identity with SEQ ID NO: 2), or the GG genotype or GX haplotype when referring to the complementary strand.

[0143] - The GG genotype or GX haplotype of SNP_03 at nucleotide 81 of SEQ ID NO: 3 (or an equivalent nucleotide in a sequence having at least 95%, 96%, 97%, 98%, or 99% sequence identity with SEQ ID NO: 3).

[0144] - The AA genotype or AX haplotype of SNP_04 at nucleotide 101 of SEQ ID NO: 4 (or an equivalent nucleotide in a sequence having at least 95%, 96%, 97%, 98%, or 99% sequence identity with SEQ ID NO: 4), or the TT genotype or TX haplotype when referring to the complementary strand.

[0145] - The TT genotype or TX haplotype of SNP_05 at nucleotide 101 of SEQ ID NO: 5 (or an equivalent nucleotide in a sequence having at least 95%, 96%, 97%, 98%, or 99% sequence identity with SEQ ID NO: 5), or the AA genotype or AX haplotype when referring to the complementary strand.

[0146] - The AA genotype or AX haplotype of SNP_06 at nucleotide 101 of SEQ ID NO: 6 (or an equivalent nucleotide in a sequence having at least 95%, 96%, 97%, 98%, or 99% sequence identity with SEQ ID NO: 6), or the TT genotype or TX haplotype when referring to the complementary strand, and / or

[0147] - The CC genotype or CX haplotype of SNP_01 at nucleotide 81 of SEQ ID NO: 7 (or an equivalent nucleotide in a sequence having at least 95%, 96%, 97%, 98% or 99% sequence identity with SEQ ID NO: 7).

[0148] Therefore, a homozygous SNP genotype (e.g., CC) means that the donor nucleotide is present on both chromosomes 5 in the melon genome. A heterozygous SNP haplotype (e.g., CX) means that one of the two chromosomes 5 contains the donor SNP, while the other chromosome 5 may contain the recurrent parental SNP nucleotide. For example, X can be any nucleotide, such as any nucleotide other than the donor nucleotide (e.g., in this example, X could be nucleotides T, A, or G).

[0149] In one respect, the infiltrated fragment does not contain a gene encoding the protein of SEQ ID NO: 9 (TIR-NBS-LRR protein of QTL5) or SEQ ID NO: 10 (susceptible TIR-NBS-LRR protein). Therefore, in one respect, the plant or plant cell does not contain a gene encoding the protein of SEQ ID NO: 9 (TIR-NBS-LRR protein of QTL5) or SEQ ID NO: 10 (susceptible TIR-NBS-LRR protein). Of course, when QTL5rb exists in a heterozygous form in a plant, another chromosome 5 may contain, for example, SEQ ID NO: 9 or SEQ ID NO: 10.

[0150] In one respect, the donor plant is a species of melon, such as the wild plant of the melon subspecies.

[0151] In one respect, the donor plant of the present invention is not the short-haired melon subspecies.

[0152] In one aspect, the donor plant of the present invention is not one of the five ToLCNDV-resistant subspecies of *Momordica momordica* identified by Lopez et al., 2015, Euphytica 204(3), 679-691 (Momordica momordica varieties: Mom-KhaInd / Kharbuja, Mom-PI124Ind / PI124112, Mom-PI124Ind / PI414723 and wild-type Momordica momordica: Ag-WM9Ind / WM9, Ag-WM7Ind / WM7).

[0153] The term “breeding” in this article encompasses hybridization, backcrossing, self-crossing, selection, double haploid production, embryo rescue, protoplast fusion, marker-assisted selection, and mutation breeding, all of which are techniques known to breeders (i.e., methods other than genetic modification / transformation / transgenic methods) and can be used to obtain, identify, generate, and / or transfer recombinant chromosome 5, for example.

[0154] In a preferred embodiment of the invention, the infiltrated fragment is derived from wild melon plants or wild melon germplasm, and most preferably from a wild hairy melon subspecies (e.g., with fruit having sour pulp). This donor is used in the present invention and has been deposited with accession number NCIMB 44139. However, those skilled in the art can identify other donors that contain, for example, the same SNP genotype or SNP haplotype as the donor for one or more of SNPs 01 to SNP 07, or SNP 02 to SNP 05, or SNP 02 to SNP 06, or SNP 03 to SNP 05, or SNP 03 and / or SNP 04, and / or contain a gene encoding the TIR-NBS-LRR protein of SEQ ID NO: 8, or a protein encoding a protein having at least 96%, 97%, 98%, 99% or higher sequence identity with the protein of SEQ ID NO: 8, and said protein preferably contains the above-described and, as described above, compared to the susceptible TIR-NBS-LRR protein of SEQ ID NO: 10 (encoded by a gene present in the susceptible reference genome). Figure 2 The changes numbered 1, 2, 3, and 4 represent one or more amino acid changes in the amino acid changes.

[0155] In one respect, suitable donors are wild melon plants or germplasm with a mean ToLCNDV-ES disease score (preferably using a strain that destroys resistance) of at least 7.7, 7.8, 7.9, 8.0 or 8.5 or 8.6, 8.7, 8.8, 8.9 or 9.0 on a scale from 1 = plant death to 9 = asymptomatic. Such donors also preferably contain, for example, the same SNP genotype or SNP haplotype present in NCIMB44139 for one or more or all of SNP_01 to SNP_07, or SNP_02 to SNP_05, or SNP_02 to SNP_06, SNP_03 to SNP_05, or SNP_03 and / or SNP_04, and / or contain a gene encoding the protein of SEQ ID NO: 8, or a protein encoding a protein having at least 96%, 97%, 98%, 99% or higher sequence identity with the TIR-NBS-LRR resistance protein of SEQ ID NO: 8, and said protein preferably contains, as described above, the susceptible TIR-NBS-LRR protein of SEQ ID NO: 10 (encoded by a gene present in the susceptible reference genome), compared to the susceptible TIR-NBS-LRR protein of SEQ ID NO: 10. Figure 2 The changes numbered 1, 2, 3, and 4 represent one or more amino acid changes in the amino acid changes.

[0156] In one particular embodiment, the plant cells and plants provided herein are characterized in that the introgression fragments or QTL5rb or the potential causal gene of QTL5rb (conferring ToLCNDV-ES resistance) present in the cells are derived from seeds preserved with NCIMB 44139 or their progeny, whereby the progeny retain the gene encoding the TIR-NBS-LRR resistance protein of SEQ ID NO: 8.

[0157] Therefore, in one aspect, a melon plant cell or melon plant contains the genome sequence of SEQ ID NO: 11 on chromosome 5 in its genome, which encodes the protein of SEQ ID NO: 8. The plant cell or plant will transcribe the mRNA (described as cDNA) of SEQ ID NO: 12. In one aspect, the melon plant or plant cell contains SEQ ID NO: 11, but has some nucleotide variations in the intron sequence of SEQ ID NO: 11. In one aspect, the melon plant or plant cell contains a genome sequence wherein the exon sequence is preferably 100% identical to the exon sequence of SEQ ID NO: 11. The exon sequence can be identified by comparing SEQ ID NO: 11 with the cDNA (mRNA) of SEQ ID NO: 12. Therefore, in one aspect, a melon plant cell or melon plant contains the genome sequence encoding the protein of SEQ ID NO: 8 on chromosome 5 in its genome. Therefore, in one respect, melon plant cells or melon plants contain a genomic sequence on chromosome 5 in their genome that will transcribe the mRNA (described as cDNA) of SEQ ID NO: 12.

[0158] For example, the genome sequence of the susceptible reference genome DHL92 was found to be identical to that of the susceptible variety Vedrantais, except that the third intron in the Vedrantais sequence is missing a single nucleotide, i.e., it has a one-nucleotide gap. These two genome sequences encode the same susceptibility protein as SEQ ID NO: 10 (as discovered using Needle in protein pairing, thus the sequence identity percentage between SEQ ID NO: 10 and the susceptibility protein sequence of Vedrantais is 100%).

[0159] In another preferred embodiment, the provided melon plant cells are derived from a cultivated melon plant, or the melon plant provided herein is a cultivated melon plant, and the infiltrated fragment or QTL5rb is derived from wild melon germplasm or from wild long-haired melon subspecies germplasm, or from a donor plant described herein as a preferred donor plant, or obtained from / available from seeds or their progeny preserved in NCIMB 44139.

[0160] In another aspect, melon plants and melon plant cells are provided in which an endogenous gene encoding a susceptible TIR-NBS-LRR protein on chromosome 5 (which does not confer resistance) has been genetically edited or mutated to encode a TIR-NBS-LRR protein that confers resistance to the destructive ToLCNDV-ES strain. The susceptible plants and cells contain a gene encoding the protein of SEQ ID NO: 10, or a protein having at least 98%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.7%, 98.8%, 98.9%, 99%, 99.5%, or 99.8% sequence identity with the protein of SEQ ID NO: 10. The gene encoding this "susceptibility protein" can be edited to encode a "resistance protein" by modifying one, two, three, or all of the four amino acid differences in the "susceptibility protein," which confers resistance to the protein, i.e., transforms it into a "resistance protein."

[0161] As mentioned, the introgression fragment conferring resistance to QTL5rb comprises a gene encoding the protein of SEQ ID NO: 8 or a protein having at least 96%, 97%, 98%, or 99% sequence identity with the protein of SEQ ID NO: 8, and said protein preferably contains one or more of the following four amino acid changes compared to the protein of SEQ ID NO: 10:

[0162] - "Change 1": At the beginning of the protein, two S (serine) residues are inserted after amino acid E9 (glutamic acid 9); that is, the protein contains 9 serine residues (S) after E9, and amino acids 10 to 18 are serine residues in proteins that have at least 96%, 97%, 98%, or 99% sequence identity with the protein of SEQ ID NO: 8 (while SEQ ID NO: 10 contains only 7 serine residues, i.e., amino acids 10 to 16); see also Figure 2 Change number 1;

[0163] - "Change No. 2": At the equivalent position in a protein having at least 96%, 97%, 98%, or 99% sequence identity with the protein of SEQ ID NO: 8, amino acid V483 (valine 483 of SEQ ID NO: 10) is changed to M (methionine); in SEQ ID NO: 8, the equivalent position is M484; see also Figure 2 Change number 2;

[0164] - "Change No. 3": Amino acid E628 (glutamic acid 628 of SEQ ID NO: 10) is changed to K (lysine) at the equivalent position in proteins having at least 96%, 97%, 98%, or 99% sequence identity with the protein of SEQ ID NO: 8; in SEQ ID NO: 8, the equivalent position is K629; see also Figure 2 Change number 3;

[0165] - "Change No. 4": At the equivalent position in a protein having at least 96%, 97%, 98%, or 99% sequence identity with the protein of SEQ ID NO: 8, amino acid Y678 (tyrosine 678 of SEQ ID NO: 10) is changed to H (histidine); in SEQ ID NO: 8, the equivalent position is H679, see [link to relevant documentation]. Figure 2 Change 4; or at the equivalent position in a protein having at least 96%, 97%, 98%, or 99% sequence identity with the protein of SEQ ID NO: 8, amino acid Y678 (tyrosine 678 of SEQ ID NO: 10) is changed to D (aspartic acid); in SEQ ID NO: 8, the equivalent position is D679, see [link to SEQ ID NO: 8]. Figure 1 or Figure 2 Change number 4.

[0166] For example, introducing one or more of these four types of changes into the susceptible protein through gene editing technology (such as CRISPR-based technology) or random mutagenesis technology (such as chemical mutagenesis or radiation mutagenesis) will transform the susceptible protein into a resistant protein.

[0167] In one aspect, the gene encoding the susceptibility protein of SEQ ID NO: 10 (or a protein having at least 98%, 98.5%, 99%, 99.5%, or 99.8% sequence identity with SEQ ID NO: 10) is modified by introducing one or more of the four variations (variations 1 to 4) into the gene and the encoded protein. In SEQ ID NO: 36, a genomic sequence encoding the susceptibility protein of SEQ ID NO: 10 on chromosome 5 is provided. Therefore, in one aspect, the genomic gene sequence of SEQ ID NO: 36 or the genomic sequence encoding the susceptibility protein having at least 98%, 98.5%, 99%, 99.5%, or 99.8% sequence identity with SEQ ID NO: 10 is modified, for example, by producing one or more of the variations 1, 2, 3, and / or 4 described above.

[0168] Figure 4The diagram illustrates four potential codon variations. A “susceptible protein” can be modified to a “resistant protein” by, for example, modifying one or more codons in the genome sequence of, for example, SEQ ID NO: 36 or the genome sequence encoding a susceptible protein having at least 98%, 98.5%, 99%, 99.5%, or 99.8% sequence identity with the susceptible protein in SEQ ID NO: 10 (e.g., inserting one or more codons, changing codons to encode different amino acids, etc.).

[0169] In one aspect, a modified or mutated melon plant, plant cell, or plant part is provided, wherein the plant, plant cell, or plant part contains an endogenous TIR-NBS-LRR gene at a locus on chromosome 5, wherein the endogenous gene is modified or mutated to encode a resistance protein, such as the protein of SEQ ID NO: 10 (or a protein having at least 98%, 98.5%, 99%, 99.5%, or 99.8% sequence identity with SEQ ID NO: 10), modified to include one or more variations of SEQ ID NO: 1, 2, 3, and / or 4, or wherein the endogenous gene is modified or mutated to encode a resistance protein having at least 96%, 97%, 98%, or 99% sequence identity with the resistance protein of SEQ ID NO: 8 and including one or more variations of SEQ ID NO: 1, 2, 3, and / or 4. Attached Figure Description

[0170] Figure 1 Multiple sequence alignments of the TIR-NBS-LRR protein from SEQ ID NO: 8 (QTL5rb), SEQ ID NO: 9 (QTL5 of US11,591,611), and SEQ ID NO: 10 (reference genomes of susceptible varieties Vedrantais and DHL92v4). Dashed boxes indicate leucine-rich repetitive sequences present in the proteins.

[0171] Figure 2 Paired sequence alignment of SEQ ID NO: 8 (resistance protein) and SEQ ID NO: 10 (susceptibility protein), where the four distinct variations between the resistance and susceptibility proteins are indicated by numbers 1 (variation 1), 2 (variation 2), 3 (variation 3), and 4 (variation 4). These are also... Figure 1 As shown in the image.

[0172] Figure 3 SNP haplotypes of QTL5 in QTL5rb and US11,591,611; the SNP nucleotides shown are nucleotides present on the positive strand relative to the reference genome.

[0173] Figure 4Paired sequence alignments of the genomic sequences of the susceptibility protein (SEQ ID NO: 36) encoding SEQ ID NO: 10 and the resistance protein (SEQ ID NO: 11) encoding SEQ ID NO: 8. Differences at the nucleotide level are shown in boxes, resulting in four protein-level differences (numbered 1 to 4, i.e., changes 1 through 4). For example, change 1 involves inserting the codon TCT encoding serine twice, resulting in two serine residues in the resistance protein. Change 2 is the change of codon GTG (valine, V) to ATG (methionine, M), change 3 is the change of codon GAG (glutamate, E) to AAG (lysine, K), and change 4 is the change of codon TAT (tyrosine, Y) to CAT (histidine, H).

[0174] Figure 5 The mean disease score of 15 plants (N=15) of each genotype at 40 dpi (days after inoculation or infection) is shown on a scale from 1 (plant death) to 9 (asymptomatic), as described in the examples.

[0175] Figure 6 The figure shows the average ct value (q-PCR) of 10 plants (N=10) for each genotype at 55 dpi.

[0176] General definition

[0177] As used herein, the term "plant" includes the whole plant or any part or derivative thereof (preferably having the same genetic composition as the plant from which it is obtained), such as plant organs (e.g., harvested or unharvested fruits, leaves, flowers, anthers, etc.), plant cells, plant protoplasts, plant cell tissue cultures capable of regenerating a whole plant, plant callus, plant cell masses, plant grafts, seedlings, whole plant cells in a plant, plant clonal or micropropagation, or plant parts such as plant cuttings, embryos, pollen, anthers, ovules, fruits (e.g., harvested tissues or organs), flowers, leaves, seeds, cloned plants, roots, stems, root tips, grafts (scions and / or rootstocks), etc. It also includes any developmental stage, such as seedlings, pre- or post-rooted cuttings, etc. When referring to "seeds of a plant," these are seeds that a plant can produce from its own seeds or from the plant after self-fertilization or cross-fertilization.

[0178] As used herein, the term "variety" or "cultivar" refers to a group of plants within a single botanical taxa at the lowest known rank, which can be defined by the expression of characteristics produced by a given genotype or combination of genotypes. A plant characterized by the presence of an allele of a single gene is not a variety or cultivar because the rest of its genome has not been characterized.

[0179] "F1, F2, F3, etc." refers to the consecutive generations following a cross between two parent plants or parent strains. Plants grown from seeds produced by crossing two plants or strains are called the F1 generation. F1 plants self-pollinate to produce the F2 generation, and so on.

[0180] "F1 hybrid" plants (or F1 hybrid seeds) are the generation obtained by crossing two inbred parent lines. Therefore, F1 hybrid seeds are the seeds from which F1 hybrid plants grow. Due to heterosis, F1 hybrids are more vigorous and have higher yields. Inbred lines are essentially homozygous at most loci in their genome.

[0181] “Plant strain” or “breeding strain” refers to a plant and its offspring. As used herein, the term “inbred strain” refers to a plant strain that has undergone repeated self-pollination and is nearly homozygous. Therefore, “inbred strain” or “parental strain” refers to a plant that has undergone several generations (e.g., at least 5, 6, 7 or more generations) of close inbreeding, resulting in a plant strain with a high degree of uniformity.

[0182] "Conformity" or "uniformity" refers to the genetic and phenotypic characteristics of a plant line or variety. Inbred lines are genetically highly consistent because they are produced through several generations of close inbreeding. Similarly, the F1 hybrids produced from such inbred lines are highly consistent in their genotype and phenotypic characteristics and performance.

[0183] In the context of this invention, "chromosome 5 of a melon plant" should be understood to mean, for example, chromosome 5 found on the melonomics.net website, DHL92 version 4.

[0184] "Orthologous chromosome 5" refers to chromosome 5 of wild melons or wild relatives of melons, a portion of which can infiltrate into chromosome 5 of cultivated melons.

[0185] "Recombinant chromosome 5" refers to chromosome 5 having a new genetic composition resulting from the exchange between homologous chromosomes. For example, "recombinant chromosome 5" is chromosome 5 that is not present in either parent plant and is produced through a rare exchange event between homologous chromosomes of chromosome 5 pairs. In this document, for example, a recombinant melon chromosome 5 containing a QTL5rb conferring resistance to ToLCNDV-ES is provided. Thus, in one aspect, recombinant chromosome 5 is a chromosome of cultivated melon with an introgression segment from a wild donor, whereby the introgression segment contains a QTL5rb conferring resistance to ToLCNDV-ES.

[0186] "QTL5rb potential causal gene" in this document refers to the TIR-NBS-LRR gene encoding the resistance protein of SEQ ID NO: 8 or a protein having at least 96% sequence identity with SEQ ID NO: 8 and preferably containing at least one of the following amino acids, preferably at least two, three, or all four amino acids:

[0187] - Nine serines (S) at equivalent positions in amino acids 10 to 18 of SEQ ID NO: 8, or in a sequence having at least 96% sequence identity with SEQ ID NO: 8.

[0188] - Methionine (M) at amino acid number 484 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0189] - Lysine (K) at amino acid number 629 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0190] - Histidine (H) or aspartic acid (D) at amino acid number 679 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8.

[0191] In one respect, the potential causal gene of QTL5rb can be identified in the donor plant and infiltrated into cultivated melons, or it can be generated de novo, for example, by targeted gene editing or by random mutagenesis modifying the endogenous TIR-NBS-LRR gene (encoding, for example, a susceptibility protein) at the locus.

[0192] "Modified TIR-NBS-LRR gene" or "QTL5rb potential modified causal gene" refers to chromosome 5 in which the endogenous TIR-NBS-LRR gene has been modified, for example, by targeted gene editing or by random mutagenesis, such that the protein encoded by the modified gene is the protein of SEQ ID NO: 8, or a protein having at least 96%, 97%, 98%, 99% or higher sequence identity with the protein of SEQ ID NO: 8, and said protein preferably includes, compared to the protein of SEQ ID NO: 10 (encoded by a gene present in a susceptible reference genome), the protein described elsewhere herein and as such. Figure 2 One or more amino acid changes are shown in the amino acid changes indicated by changes numbered 1, 2, 3 and 4.

[0193] “Targeted gene editing” refers to techniques that can modify endogenous target genes, such as inserting, replacing, and / or deleting one or more nucleotides in, for example, promoters or coding sequences. For example, CRISPR-based techniques, such as CRISpr-Cas9 gene editing, CRISpr-CpfI gene editing, or the more recent techniques known as “base editing” or “primer editing,” can be used to modify endogenous target genes, such as the endogenous TIR-NBS-LRR gene in melon (e.g., encoding the susceptibility protein of SEQ ID NO: 10, or the wild-type susceptibility protein having at least 98%, 98.3%, 98.4%, 98.5%, 98.6%, 98.7%, 98.8%, or 98.9% sequence identity with SEQ ID NO: 10).

[0194] "Random mutagenesis" refers to a method that introduces mutations at random locations in the genome, thereby allowing selection of mutations in target genes. Random mutagenesis includes, for example, treating seeds or plant cells or parts with mutagens such as radiation (e.g., UV treatment) or chemical mutagens such as EMS.

[0195] "DH plants," or "double haploid plants," are diploid plants produced by doubling the haploid genome of diploid plants using techniques such as in vitro methods. Therefore, DH plants are homozygous at all loci.

[0196] "Cultivated melon plants" refers to melon plants, namely varieties, bred lines, or cultivars of the melon species, cultivated by humans and possessing good agronomic characteristics, especially producing edible and marketable fruits of good size, quality, and uniformity. These plants are not "wild melon plants," which typically have much lower yields and poorer agronomic characteristics than cultivated plants and grow naturally, for example, in wild populations. "Wild plants" include, for example, an ecotype of a species, plant introduction (PI) lines, endemic varieties, or wild germplasm or wild relatives.

[0197] In the context of this invention, "SNP (=Single Nucleotide Polymorphism)" should be understood as a single nucleotide variation occurring at a specific location in the genome. An SNP is a single nucleotide variation at a given location in the genome of two plants. If a wild melon plant with ToLCNDV-ES resistance (the donor plant) shows a nucleotide at a specific single location in its corresponding sequence that differs from the corresponding nucleotide at the same location in, for example, a cultivated melon plant, then that location defines an SNP between the wild and cultivated melons. If the donor plant has one of four possible nucleotides (A, C, T, or G) at a specific location, then an SNP occurs when, for example, the cultivated plant has any of the other three possible nucleotides at the same location in its corresponding sequence. In cultivated melon plants containing introgression fragments from the donor, it is therefore easy to determine whether the single nucleotide of the SNP originates from the donor or the cultivated melon (the recipient).

[0198] “SNP nucleotide” refers to a single nucleotide, while “SNP genotype” refers to a nucleotide pair in a diploid plant cell. Therefore, for SNP_01, the SNP nucleotide of the ToLCNDV-ES resistance donor is cytosine (C) of nucleotide 81 of SEQ ID NO: 1, and the SNP genotype of the plant or cell containing SEQ ID NO: 1 can be “CC” (cytosine on both chromosomes) or “CX” (cytosine on one chromosome 5 and X on the other chromosome 5, i.e., a different nucleotide, such as thymine; or this can also be written as “C”), thus the ToLCNDV-ES resistance donor SNP nucleotide (cytosine), and therefore SEQ ID NO: 1 (or a sequence substantially the same as SEQ ID NO: 1) is homozygous or heterozygous. "SNP haplotype" refers to the nucleotide marker of an SNP on chromosome 5 in a diploid cell. Therefore, the SNP haplotype of SNP_01–SNP_02–SNP_03–SNP_04 is CGGT, see example. Figure 3 .

[0199] The verb "include" and its inflections are used in a non-restrictive sense, meaning to include the items that follow the word, but not excluding items not specifically mentioned. Furthermore, the indefinite article "a" or "one / kind" does not preclude the possibility of more than one / kind of an element, unless the context clearly requires the presence of one and only one / kind of an element. Therefore, the indefinite article "a" or "one / kind" usually means "at least one / kind," for example, "plant" also refers to several cellular plants, etc. Similarly, "fruit" or "plant" also refers to several fruits and plants.

[0200] The term "allelic gene" means any one or more alternative forms of a gene at a specific locus, such as the TIR-NBS-LRR locus (where the TIR-NBS-LRR gene is located on chromosome 5; on melonomics.net V4, the genomic transcription region of the TIR-NBS-LRR gene is located at nucleotide 26.034.368 and terminates at nucleotide 26.027.838 on chromosome 5); the allele of a gene can be an allele encoding a susceptible TIR-NBS-LRR protein that does not confer resistance to ToLCNDV-ES (e.g., the protein of SEQ ID NO: 10) (also referred to as a "susceptible protein") or an allele encoding a resistant TIR-NBS-LRR protein that confers resistance to ToLCNDV-ES (e.g., the protein of SEQ ID NO: 8 or a protein having at least 96% sequence identity with SEQ ID NO: 8) (also referred to as a "resistant protein"). In the diploid cells of an organism, the alleles of a given gene are located at specific locations or loci (multiple loci) on a chromosome. An allele exists on each chromosome of a homologous chromosome pair. Diploid plant species can contain a large number of different alleles at a given locus. These can be the same allele of the gene (homozygous) or two different alleles (heterozygous).

[0201] The “TIR-NBS-LRR gene” is a single gene in melon identified on chromosome 5 that encodes a “resistance protein” (e.g., the protein of SEQ ID NO: 8 or a protein with at least 96% sequence identity to SEQ ID NO: 8) or a “susceptibility protein” (e.g., the protein of SEQ ID NO: 10). This gene is contained in a transcriptional region on a reference genome found, for example, melonomics.net V4, starting at nucleotide 26.034.368 and ending at nucleotide 26.027.838 on chromosome 5. A single copy (i.e., heterozygous) of the allele encoding the “resistance protein” in the plant genome is sufficient to confer a resistance phenotype in the plant.

[0202] The term "gene" refers to a (genomic) DNA sequence that comprises a region (transcriptional region) that is transcribed into pre-mRNA, processed in the cell (by intron splicing) into messenger RNA molecules (mRNA), and a regulatory region (e.g., promoter) that is operatively linked. An example is the TIR-NBS-LRR gene described herein. Thus, different alleles of a gene are different alternative forms of that gene, which can take the form of, for example, differences in one or more nucleotides in the genomic DNA sequence (e.g., promoter sequence, exon sequence, intron sequence, etc.), the amino acid sequence of the mRNA, and / or the protein it encodes.

[0203] “Induced mutant alleles” are mutant alleles that are induced by / have been induced by human intervention, such as by physical or chemical mutagenesis or by mutagenesis, such as by tissue culture (as described in Zhang et al., PLOS9(5) e96879) (including targeted gene editing techniques such as CRISPR-based techniques, TALENS, base editing, etc.) and insertion mutagenesis techniques such as targeted transposon insertion techniques, etc.) (for the review, see Gao et al. https: / / doi.org / 10.1016 / j.cell.2021.01.005).

[0204] "Vegetative propagation" or "clonal propagation" refers to the propagation of plants from vegetative tissues, for example, through in vitro propagation or grafting methods (using scions and rootstocks). In vitro propagation involves in vitro cell or tissue culture and the regeneration of an entire plant from in vitro culture. Grafting involves propagating the original plant by grafting it onto a rootstock. Therefore, clones of the original plant (i.e., genetically identical vegetative propagation) can be generated through in vitro culture or grafting. "Cell culture" or "tissue culture" refers to the in vitro culture of plant cells or tissues. "Regeneration" refers to the development of a plant from cell culture, tissue culture, or vegetative propagation. "Non-propagating cells" refer to cells that cannot regenerate a complete plant.

[0205] The terms "melon plant cell," "melon plant," or "cultivated melon plant or cell," also referred to in this art as muskmelon plant cell or muskmelon plant, should be understood in the context of this invention as plant cells derived from the species *Melastoma cantaloupe* or plants belonging to the species *Melastoma cantaloupe*. Melons can be classified as: Roman melon, smooth-skinned melon, and netted melon. Roman melon, also known as cantaloupe, is primarily round, with prominent ribs and almost no netting. Most have sweet, orange flesh and are typically very fragrant. North American 'cantaloupe' is not of this type, but rather belongs to the true muskmelon category, unlike European cantaloupes. Smooth-skinned melons (or winter melons) can be further subdivided into different types, such as honeydew melon, Spanish melon, sugar melon, Japanese melon, etc. Netted melons are true muskmelons with a netted rind (netted structure) and include Gallia melon, Salle melon, and North American cantaloupe.

[0206] Cultivated melons and their wild relatives are diploid and have 12 pairs of homologous chromosomes, numbered 1 to 12.

[0207] "Cultivated melon plants" refers to melon plants, namely varieties, bred lines, or cultivars of the species melon, cultivated by humans and possessing good agronomic characteristics, especially producing edible and marketable fruits of good size, quality, and uniformity; preferably, such plants are not "wild melon plants," i.e., plants that generally have much lower yields and worse agronomic characteristics than cultivated plants and grow naturally, for example, in wild populations. "Wild melon plants" include, for example, an ecotype of a species, a plant introduction (PI) line, a local variety, or wild germplasm or wild related species.

[0208] "SNP marker" refers to, for example, a single nucleotide polymorphism between the resistance TIR-NBS-LRR allele and the susceptibility TIR-NBS-LRR allele, or one or more SNP markers linked to the resistance TIR-NBS-LRR allele, such as... Figure 3 As shown. SNP marker assays (e.g., in allele-specific assays) that can distinguish between resistance and susceptibility alleles of the TIR-NBS-LRR gene can be used to screen plants, plant parts, or DNA derived therefrom for the presence of a resistance allele, such as the protein encoding SEQ ID NO: 8. For any SNP marker, an SNP marker assay can be designed based on the sequence provided herein. Such SNP marker assays can be used to detect resistance alleles, for example, in marker-assisted selection and / or SNP genotyping assays. Therefore, SNP marker assays can be used to screen plants, plant parts, or DNA derived therefrom for the presence of resistance alleles.

[0209] "INDEL markers" refer to insertion / deletion polymorphisms, such as those between resistance TIR-NBS-LRR alleles and susceptibility alleles. INDEL marker assays (e.g., allele-specific assays) can be used to screen plants, plant parts, or DNA derived from them based on the presence of resistance alleles. For example, variation 1 is caused by INDEL; see [link to INDEL assay]. Figure 4 .

[0210] Genotyping is a method that determines the genotype or allelic composition of a plant, plant part, or seed. Biallelic genotyping assays (such as KASP assays) can distinguish between two alleles at a locus. For example, SNP markers or INDEL markers can be analyzed in genotyping assays.

[0211] "Chromosomal region containing the resistance TIR-NBS-LRR allele" refers to a genomic region of chromosome 5, for example, in cultivated melons, that carries the resistance allele. The presence of the allele can be determined phenotypically and / or by the presence of one or more molecular markers linked to the resistance allele, such as SNP markers, INDEL markers, or other markers, or preferably by markers that distinguish different TIR-NBS-LRR alleles, or by the genomic sequence of the allele sequence itself. If a marker is physically coupled to the resistance TIR-NBS-LRR allele, it is "linked to that allele." "Allele-specific markers" are markers that are specific to a particular allele (e.g., a specific resistance allele) and thus distinguish, for example, the resistance allele from other alleles. Allele-specific markers are preferably markers within the allele itself, i.e., in the promoter region or transcription region of the gene, for example based on polymorphism between the resistance allele sequence and the susceptibility allele sequence. For example, the variations 1, 2, 3 and / or 4 described herein can be used to design SNP markers or INDEL markers or other markers for genotyping and for detecting alleles present in the genome.

[0212] A pair of "flank markers" refers to two markers, preferably two SNP markers or two sequences containing SNP markers, that are linked to a resistance allele such as QTL5rb, whereby the resistance allele is located between the two markers or between two sequences containing these markers.

[0213] "Brix (degree Brix / ° brix)" refers to the average total soluble solids content measured using a refractometer on several ripe fruits. Preferably, the average value is calculated for at least three types of fruits, each measured between the center and the outer skin of an open fruit.

[0214] The “physical distance” between loci on the same chromosome (e.g., between molecular markers and / or between phenotypic markers) is the actual distance expressed in bases or base pairs (bp), kilobases or kilobase pairs (kb), or megabases or megabase pairs (Mb).

[0215] The “genetic distance” between loci on the same chromosome (e.g., between molecular markers and / or phenotypic markers) is measured by the frequency of recombination, or recombination frequency (RF), and is indicated in centimoles (cM). One centimeter corresponds to approximately 1% recombination frequency. If no recombinants are found, the RF is zero, and the loci are physically extremely close together or they are identical. The farther apart two loci are, the higher the RF.

[0216] "Conformity" or "uniformity" refers to the genetic and phenotypic characteristics of a plant line or variety. Inbred lines are genetically highly consistent because they are produced through several generations of close inbreeding. Similarly, the F1 hybrids produced from such inbred lines are highly consistent in their genotype and phenotypic characteristics and performance.

[0217] If a genetic element, introgression, gene, or allele that confers a trait (such as ToLCNDV-ES resistance) is transferred from a plant or seed in which it exists to another plant or seed (such as a ToLCNDV-ES susceptible line or variety) in which it does not exist, using conventional breeding techniques, and does not result in a change in the phenotype of the recipient plant other than the addition of the trait conferred by the genetic element, locus, introgression, gene, or allele, then the genetic element, introgression, gene, or allele is said to be "obtainable from" or "possibly obtainable from" or "derived from" or "possibly derived from" or "present in" or "found in" a plant, seed, tissue, or cell. These terms are used interchangeably, and therefore a genetic element, locus, introgression, gene, or allele can be transferred to any other genetic background lacking that trait. Cultivated melons containing genetic elements, loci, introgression fragments, genes, or alleles (e.g., resistance TIR-NBS-LRR alleles) can be de novo produced, for example, by mutagenesis (e.g., chemical mutagenesis, CRISPR-Cas induction, etc.) and then hybridized, for example, with other cultivated melons.

[0218] The term “mean” or “average” in this document refers to the arithmetic mean, and the two terms are used interchangeably. Therefore, the term “mean” or “average” refers to the arithmetic mean of several measurements. Those skilled in the art will understand that the phenotype of a plant line or variety depends to some extent on growing conditions, and therefore it is preferable to measure the arithmetic mean of at least 8, 9, 10, 11, 12, 13, 14, 15, 20, 30, 40, 50, or more plants (or plant parts) in a randomized experimental design with several replicates and in suitable control plants grown under the same conditions in the same experiment. “Statistically significant” or “statistically significantly” different or “significantly” different means that a characteristic of a plant line or variety, when compared with a suitable control, shows a statistically significant difference from the control (mean) (e.g., a p-value less than 0.05 using ANOVA, p < 0.05).

[0219] The term “traditional breeding techniques” in this article encompasses hybridization, backcrossing, self-crossing, selection, double haploid production, chromosome doubling, embryo rescue, protoplast fusion, marker-assisted selection, and mutation breeding, all of which are known to breeders (i.e., methods other than transformation / transgenic methods). These techniques, for example, allow the acquisition, identification, and transfer of chromosome 5 containing the resistance TIR-NBS-LRR allele.

[0220] "Backcrossing" refers to a breeding method in which a (single) trait (such as ToLCNDV-ES resistance) can be transferred from one (usually a poorer) genetic background (also called a "donor") to another (usually a better) genetic background (also called a "recurrent parent"). The offspring of the hybridization (e.g., F1 plants obtained by crossing, for example, a donor with a recurrent parent of melon, or F2 or F3 plants obtained by self-pollination of F1, etc.) are "backcrossed" with a parent having, for example, a better genetic background. After repeated backcrossing, the trait from one (usually a poorer) genetic background is incorporated into another (usually a better) genetic background.

[0221] "Marker-assisted selection" or "MAS" is the process of selecting plants for the presence of a specific genome or region or allele using the presence of a molecular marker (such as an SNP marker or INDEL marker) that is genetically and physically associated with a specific locus or chromosomal region or allele-specific marker. For example, a molecular marker that is genetically and physically linked to the resistance TIR-NBS-LRR allele or an allele-specific marker can be used to detect and / or select, for example, melon plants or plant parts containing a resistance allele. The closer the linkage of a molecular marker to a locus, the lower the likelihood of the marker dissociating from the locus through meiotic recombination. Similarly, the closer two markers are associated with each other, the lower the likelihood of the two markers separating from each other (and the higher the likelihood of them co-segregating as a unit). Allele-specific markers are preferred because they directly target alleles for selection. For example, variations 1, 2, 3, and / or 4 can be used to develop markers that distinguish the presence of different alleles.

[0222] A molecular marker (or sequence containing a molecular marker) that differs from another marker (or sequence containing a molecular marker) or locus by 5 Mb, 3 Mb, 2.5 Mb, 2 Mb, 1 Mb, 0.5 Mb, 0.4 Mb, 0.3 Mb, 0.2 Mb, 0.1 Mb, 74 kb, 50 kb, 20 kb, 10 kb, 5 kb, 2 kb, 1 kb or less refers to a genomic DNA region physically located flanking (i.e., on either side of) the marker within 5 Mb, 3 Mb, 2.5 Mb, 2 Mb, 1 Mb, 0.5 Mb, 0.4 Mb, 0.3 Mb, 0.2 Mb, 0.1 Mb, 74 kb, 50 kb, 20 kb, 10 kb, 5 kb, 2 kb, 1 kb or less.

[0223] The "LOD score" (logarithm of odds, base 10) is a statistical test commonly used in linkage analysis in animal and plant populations. The LOD score compares the probability of obtaining test data with the probability of observing the same data purely by chance, assuming a genuine association between two loci (molecular marker loci and / or phenotypic trait loci). A positive LOD score favors the presence of linkage, and an LOD score greater than 3.0 is considered evidence of linkage. An LOD score of +3 indicates a 1000 to 1 probability that the observed linkage is not accidental.

[0224] "Transgenic" or "chimeric gene" refers to a genetic locus containing a DNA sequence (such as a recombinant gene encoding a protein, for example, SEQ ID NO: 8) that has been introduced into the genome of a plant through transformation (such as Agrobacterium-mediated transformation). Plants containing transgenes stably integrated into their genome are called "transgenic plants".

[0225] "Isolated nucleic acid sequence" or "isolated DNA" refers to a nucleic acid sequence that is no longer present in the natural environment in which it was isolated, such as a nucleic acid sequence in a bacterial host cell or a plant nucleus or plastid genome. When the term "sequence" is used in this document, it should be understood that molecules having such sequences refer to, for example, nucleic acid molecules.

[0226] "Host cell," "recombinant host cell," or "transformed cell" are terms referring to a new single cell (or organism) produced by the introduction of at least one nucleic acid molecule into the cell. The host cell is preferably a plant cell or a bacterial cell. The host cell may contain nucleic acids as extrachromosomal (additional) repeating molecules, or nucleic acids integrated into the nuclear or plasmonic genome of the host cell, or nucleic acids as introduced chromosomes (e.g., mini-chromosomes).

[0227] "Sequence identity" and "sequence similarity" can be determined by aligning two peptide or two nucleotide sequences using global or local alignment algorithms. Sequences can be described as "substantially identical" or "substantially similar" when optimally aligned using programs such as GAP, BESTFIT, or the Emboss program "Needle" (using default parameters, see below), sharing at least a minimum percentage of sequence identity (as further defined below). These programs use the Needleman and Wunsch global alignment algorithms to align two sequences across their entire length, maximizing the number of matches and minimizing the number of gaps. Typically, default parameters are used, where the gap generation penalty = 10 and the gap extension penalty = 0.5 (for both nucleotide and protein alignments). For nucleotides, the default scoring matrix used is DNAFULL, and for proteins, the default scoring matrix is ​​Blosum62 (Henikoff and Henikoff, 1992, PNAS 89, 10915-10919). Sequence alignment and the score for sequence identity percentage can be determined, for example, using a computer program such as EMBOSS, available on the World Wide Web at ebi.ac.uk / Tools / psa / emboss_needle / . Alternatively, sequence similarity or identity can be determined by searching databases such as FASTA, BLAST, etc., but the matches should be retrieved and aligned to compare sequence identity. If the sequence identity percentage is at least 95%, 98%, 99%, or higher, the two proteins, two protein domains, or two nucleic acid sequences have “basic sequence identity” (as determined by Emboss “needle” using default parameters (i.e., nick production penalty = 10, nick extension penalty = 0.5), using the score matrix DNAFULL for nucleic acids and Blosum62 for proteins).

[0228] When referring to a nucleic acid sequence (e.g., DNA or genomic DNA) that has “basic sequence identity” with a reference sequence or has at least 90% sequence identity with a reference sequence, such as at least 95%, 96%, 97%, 98%, 99%, 99.2%, 99.5%, or 99.9% nucleic acid sequence identity, in one embodiment, the nucleotide sequence is considered to be substantially identical to the given nucleotide sequence and can be identified using stringent hybridization conditions.

[0229] “Strong hybridization conditions” can be used to identify nucleotide sequences that are substantially identical to a given nucleotide sequence. The stringent conditions depend on the sequence and will vary under different conditions. Typically, stringent conditions are chosen to be about 5°C lower than the thermal melting point (Tm) of the specific sequence at defined ionic strengths and pH. Tm is the temperature at which 50% of the target sequence hybridizes with a perfectly matched probe (at defined ionic strengths and pH). Stringent conditions are typically chosen where the salt concentration is about 0.02 mol at pH 7 and the temperature is at least 60°C. Decreasing the salt concentration and / or increasing the temperature increases stringency. Stringent conditions for RNA-DNA hybridization (using, for example, a 100 nt probe for Northern blotting) include, for example, those involving at least one wash for 20 minutes in 0.2X SSC at 63°C, or equivalent conditions. Stringent conditions for DNA-DNA hybridization (using, for example, a 100 nt probe for Southern blotting) include, for example, those involving at least one (usually two) washes for 20 minutes in 0.2X SSC at a temperature of at least 50°C, typically about 55°C, or equivalent conditions.

[0230] In the context of this invention, "M1 generation" or "M1 plant" refers to the first generation produced directly from mutagenesis treatment. For example, plants grown from seeds treated with a mutagen are representative of the M1 generation.

[0231] In this article, "M2 generation" or "M2 plant" refers to the generation obtained from self-pollination of the M1 generation. Plants grown from seeds obtained from self-pollinated M1 plants are represented as M2 plants. M3, M4, etc., refer to other generations obtained after self-pollination.

[0232] “mRNA coding sequence” has its usual meaning in this article. Except for the substitution of thymine (T) with uracil (U), the mRNA coding sequence corresponds to the corresponding DNA coding (cDNA) sequence of the gene / allele.

[0233] A "mutation" in a nucleic acid molecule (DNA or RNA) is a change of one or more nucleotides compared to the corresponding wild-type sequence, for example, through the substitution, deletion, or insertion of one or more nucleotides. Examples of such mutations are point mutations, nonsense mutations, missense mutations, splice site mutations, frameshift mutations, or mutations in regulatory sequences.

[0234] "Nucleic acid molecule" has the common understanding in this field. It is composed of nucleotides containing either the sugar deoxyribose (DNA) or ribose (RNA).

[0235] A point mutation is a substitution of a single nucleotide, or an insertion or deletion of a single nucleotide.

[0236] A nonsense mutation is a point mutation in the nucleic acid sequence that encodes a protein, thereby changing a codon in the nucleic acid molecule to a stop codon. This results in a premature stop codon in the mRNA and leads to the translation of a truncated protein. Truncated proteins may have reduced or lost function.

[0237] "Missense or nonsynonymous mutations" are point mutations in the nucleic acid sequence that encodes a protein, thereby changing the codon to encode a different amino acid. The resulting protein may have reduced or lost its function.

[0238] "Splicing site mutation" is a mutation in the nucleic acid sequence that encodes a protein, thereby altering the RNA splicing of the pre-mRNA and producing an mRNA with a different nucleotide sequence and a protein with a different amino acid sequence than the wild type. The resulting protein may have reduced or lost function.

[0239] A frameshift mutation is a mutation in the nucleic acid sequence that encodes a protein. This mutation alters the reading frame of the mRNA, resulting in a different amino acid sequence. The resulting protein may have reduced or lost its function.

[0240] In the context of this invention, "deletion" means the absence of at least one nucleotide anywhere in a given nucleic acid sequence compared to the corresponding wild-type sequence, or the absence of at least one amino acid anywhere in a given amino acid sequence compared to the corresponding (wild-type) sequence.

[0241] "Truncation" should be understood as the deletion of at least one nucleotide at the 3' or 5' end of the nucleotide sequence compared to the corresponding wild-type sequence, or the deletion of at least one amino acid at the N-terminus or C-terminus of the protein compared to the corresponding wild-type protein sequence, but preferably at least 10, 20, 30, 40, 50, 60, 70, 80, 90, 100 or more amino acids. The 5' end is determined by the ATG codon used as the start codon in the translation of the corresponding wild-type nucleotide sequence.

[0242] "Substitution" means that at least one nucleotide in a nucleic acid sequence or an amino acid in a protein sequence is different from the corresponding wild-type nucleic acid sequence or the corresponding wild-type amino acid sequence, due to, for example, the exchange of nucleotides in the coding sequence of the corresponding protein.

[0243] "Insertion" means that the nucleic acid sequence or the amino acid sequence of a protein contains at least one additional nucleotide or amino acid compared to the corresponding wild-type nucleic acid sequence or the corresponding wild-type amino acid sequence.

[0244] In the context of this invention, "early stop codon" means that, compared to the stop codon of the corresponding wild-type coding sequence, the start codon that is closer to the 5' end exists in the coding sequence (cds).

[0245] "Mutations in regulatory sequences," such as mutations in the promoters or enhancers of a gene, are changes in one or more nucleotides compared to the wild-type sequence, for example, by substitution, deletion, or insertion of one or more nucleotides, resulting in, for example, the production of reduced or no mRNA transcripts of the gene.

[0246] "Mutation in a protein" is, for example, a change of one or more amino acid residues compared to the wild-type sequence by means of substitution, deletion, truncation or insertion of one or more amino acid residues.

[0247] In this article, a "mutant protein" is a protein whose nucleic acid sequence encoding the protein contains one or more mutations.

[0248] "Knockout" or "complete knockout" should be understood as the expression of the corresponding gene no longer being detectable.

[0249] "Olignucleotides" or "oligonucleotide primers or probes" are short, single-stranded nucleic acid polymers, for example, with a length of at least 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or more nucleotides. Oligonucleotides can be unmodified or chemically modified in various ways depending on their intended use, such as adding 5' or 3' phosphate groups to achieve linking or blocking elongation, labeling with radionuclides or fluorophores and / or quenchers for use as probes, incorporating thiols, amino groups, or other reactive moieties to achieve covalent coupling to functional molecules such as enzymes, and elongating with other linkers and spacers of different functions. DNA oligonucleotides are the most commonly used, but RNA oligonucleotides are also available. The length of oligonucleotides is usually named by adding the suffix -mer. For example, an oligonucleotide with 19 nucleotides (bases) is called 19-mer. For most applications, oligonucleotides are designed to pair bases with DNA or RNA strands. Oligonucleotides are most commonly used as primers for PCR (polymerase chain reaction). Primers are designed so that at least a portion of their sequence is complementary to the sequence targeted for amplification. The optimal primer length for complementary sequences is, for example, 18 to 22 nucleotides. The optimal primer sequences for PCR are usually determined by primer design software.

[0250] A DNA microarray is an array of microscopic dots containing DNA (usually oligonucleotides) bound to a solid support. The target can be DNA, cDNA, or cRNA. Depending on the system, hybridization of the target to a specific dot is detected by fluorescence, chemiluminescence, or colloidal silver or gold. Microarrays are used in a variety of applications, such as simultaneous measurement of large numbers of gene expressions, enabling genome-wide gene expression analysis, and genotyping studies using methods such as single nucleotide polymorphism (SNP) or InDel analysis.

[0251] "Complementary strands" refer to the two strands of a complementary sequence, and for double-stranded DNA, they can be called sense (or positive) and antisense (or negative) strands. For any sequence provided herein, only one strand of the sequence is given, but the complementary strand of the given strand is also covered herein. The complementary nucleotides of DNA are A, which is complementary to T, and G, which is complementary to C. The complementary nucleotides of RNA are A, which is complementary to U, and G, which is complementary to C.

[0252] “SNP_01” refers to nucleotide 81 of SEQ ID NO: 1, or an equivalent nucleotide in a sequence having at least 95%, 96%, 97%, 98%, or 99% sequence identity (when paired) with SEQ ID NO: 1. SNP_01 corresponds to a specific nucleotide position on chromosome 5 of the melon genome (refer to the melon reference genome DHL92 V4 in the melonnomics.net database) as listed in Table 2. “Donor nucleotide of SNP_01” refers to the ToLCNDV-ES resistance donor nucleotide of SNP_01 shown in Table 2. SEQ ID NO: 1 is the forward strand relative to the reference genome. SNP_01 is positioned on one side of the region containing QTL5rb, and SNP_07 on the other side.

[0253] “SNP_02” refers to nucleotide 242 of SEQ ID NO: 2, or an equivalent nucleotide in a sequence having at least 95%, 96%, 97%, 98%, or 99% sequence identity (when paired) with SEQ ID NO: 2. SNP_02 corresponds to a specific nucleotide position on chromosome 5 of the melon genome (refer to the melon reference genome DHL92 V4 in the melonnomics.net database) as listed in Table 2. “Donor nucleotide of SNP_02” refers to the ToLCNDV-ES resistance donor nucleotide of SNP_02 shown in Table 2. SEQ ID NO: 2 is the reverse strand relative to the reference genome. Therefore, the nucleotide of SNP_02 on the complementary strand may be referred to herein.

[0254] “SNP_03” refers to nucleotide 81 of SEQ ID NO: 3, or an equivalent nucleotide in a sequence having at least 95%, 96%, 97%, 98%, or 99% sequence identity (when paired) with SEQ ID NO: 3. SNP_03 corresponds to a specific nucleotide position on chromosome 5 of the melon genome (refer to the melon reference genome DHL92 V4 in the melonnomics.net database) as listed in Table 2. “Donor nucleotide of SNP_03” refers to the ToLCNDV-ES resistance donor nucleotide of SNP_03 shown in Table 2. SEQ ID NO: 3 is the forward strand relative to the reference genome.

[0255] “SNP_04” refers to nucleotide 101 of SEQ ID NO: 4, or an equivalent nucleotide in a sequence having at least 95%, 96%, 97%, 98%, or 99% sequence identity (when paired) with SEQ ID NO: 4. SNP_04 corresponds to a specific nucleotide position on chromosome 5 of the melon genome (refer to the melon reference genome DHL92 V4 in the melonnomics.net database) as listed in Table 2. “Donor nucleotide of SNP_04” refers to the ToLCNDV-ES resistance donor nucleotide of SNP_04 shown in Table 2. SEQ ID NO: 4 is the reverse strand relative to the reference genome. Therefore, the nucleotide of SNP_04 on the complementary strand may be referred to herein.

[0256] “SNP_05” refers to nucleotide 101 of SEQ ID NO: 5, or an equivalent nucleotide in a sequence having at least 95%, 96%, 97%, 98%, or 99% sequence identity (when paired) with SEQ ID NO: 5. SNP_05 corresponds to a specific nucleotide position on chromosome 5 of the melon genome (refer to the melon reference genome DHL92 V4 in the melonnomics.net database) as listed in Table 2. “Donor nucleotide of SNP_05” refers to the ToLCNDV-ES resistance donor nucleotide of SNP_05 shown in Table 2. SEQ ID NO: 5 is the reverse strand relative to the reference genome. Therefore, the nucleotide of SNP_05 on the complementary strand may be referred to herein.

[0257] “SNP_06” refers to nucleotide 101 of SEQ ID NO: 6, or an equivalent nucleotide in a sequence having at least 95%, 96%, 97%, 98%, or 99% sequence identity (when paired) with SEQ ID NO: 6. SNP_06 corresponds to a specific nucleotide position on chromosome 5 of the melon genome (refer to the melon reference genome DHL92 V4 in the melonnomics.net database) as listed in Table 2. “Donor nucleotide of SNP_06” refers to the ToLCNDV-ES resistance donor nucleotide of SNP_06 shown in Table 2. SEQ ID NO: 6 is the reverse strand relative to the reference genome. Therefore, the nucleotide of SNP_06 on the complementary strand may be referred to herein.

[0258] “SNP_07” refers to nucleotide 81 of SEQ ID NO: 1, or an equivalent nucleotide in a sequence having at least 95%, 96%, 97%, 98%, or 99% sequence identity (when paired) with SEQ ID NO: 7. SNP_07 corresponds to a specific nucleotide position on chromosome 5 of the melon genome (refer to the melon reference genome DHL92 V4 in the melonnomics.net database) as listed in Table 2. “Donor nucleotide of SNP_07” refers to the ToLCNDV-ES resistance donor nucleotide of SNP_07 shown in Table 2. SEQ ID NO: 7 is the forward strand relative to the reference genome. SNP_07 is positioned on one side of the region containing QTL5rb, and SNP_01 on the other side. Detailed Implementation

[0259] In one aspect, this article provides a cultivated melon plant or plant cell of a melon species, the cultivated melon plant or plant cell containing an introgression fragment from a ToLCNDV-ES resistance donor plant on chromosome 5, wherein the introgression fragment contains a quantitative trait locus called QTL5rb conferring resistance to ToLCNDV-ES, particularly resistance to strains that destroy resistance, and wherein the introgression fragment contains guanine (G) at nucleotide 81 of SEQ ID NO: 3 or at an equivalent nucleotide of a sequence having at least 95% sequence identity with SEQ ID NO: 3 and / or adenine (A) (or thymine in the complementary strand) at nucleotide 101 of SEQ ID NO: 4 or at an equivalent nucleotide of a sequence having at least 95% sequence identity with SEQ ID NO: 4.

[0260] In one aspect, this article also provides a cultivated melon plant or plant cell of a melon species, the cultivated melon plant or plant cell containing an introgression fragment from a ToLCNDV-ES resistance donor plant on chromosome 5, wherein the introgression fragment contains a quantitative trait locus called QTL5rb conferring ToLCNDV-ES resistance, particularly resistance to resistance-destroying strains, and wherein the introgression fragment contains at least one, two, three, four, or five markers selected from the group consisting of:

[0261] - Cytosine (C) (or guanine in the complementary strand) at nucleotide 242 of SEQ ID NO: 2 or at an equivalent nucleotide of a sequence having at least 95% sequence identity with SEQ ID NO: 2 (or the complementary strand);

[0262] - Guanine at nucleotide 81 of SEQ ID NO: 3 or at an equivalent nucleotide of a sequence having at least 95% sequence identity with SEQ ID NO: 3;

[0263] - Adenine (or thymine in the complementary strand) at nucleotide 101 of SEQ ID NO: 4 or at an equivalent nucleotide of a sequence having at least 95% sequence identity with SEQ ID NO: 4 (or with the complementary strand).

[0264] - Thymine (T) (or adenine in the complementary strand) at nucleotide 101 of SEQ ID NO: 5 or at an equivalent nucleotide of SNP_05 of a sequence having at least 95% sequence identity with SEQ ID NO: 5 (or with the complementary strand).

[0265] - Adenine (A) (or thymine in the complementary strand) at nucleotide 101 of SEQ ID NO: 6 or at an equivalent nucleotide of a sequence having at least 95% sequence identity with SEQ ID NO: 6 (or with the complementary strand).

[0266] In another aspect, this article provides a cultivated melon plant or plant cell of a melon species, the cultivated melon plant or plant cell containing an introgression fragment from a ToLCNDV-ES resistance donor plant on chromosome 5, wherein the introgression fragment contains a quantitative trait locus called QTL5rb conferring resistance to ToLCNDV-ES, particularly resistance to strains that destroy resistance, and wherein the introgression fragment contains guanine (G) at nucleotide 81 of SEQ ID NO: 3 or at an equivalent nucleotide of a sequence having at least 95% sequence identity with SEQ ID NO: 3 and / or adenine (A) (or thymine in the complementary strand) at nucleotide 101 of SEQ ID NO: 4 or at an equivalent nucleotide of a sequence having at least 95% sequence identity with SEQ ID NO: 4 (or with the complementary strand), and / or wherein the introgression fragment contains a gene encoding a protein of SEQ ID NO: 8, or encoding a protein of SEQ ID NO: 8. 8. A gene containing a protein having at least 96% sequence identity, wherein said protein preferably comprises at least one of the following, preferably at least two, three, or all four:

[0267] - Nine serines (S) at equivalent positions in amino acids 10 to 18 of SEQ ID NO: 8, or in a sequence having at least 96% sequence identity with SEQ ID NO: 8.

[0268] - Methionine (M) at amino acid number 484 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0269] - Lysine (K) at amino acid number 629 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0270] - Histidine (H) or aspartic acid (D) at amino acid number 679 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0271] In yet another aspect, a cultivated melon plant or plant cell of a melon species is provided, the cultivated melon plant or plant cell containing an introgression segment from a ToLCNDV-ES resistance donor plant on chromosome 5, wherein the introgression segment contains a quantitative trait locus called QTL5rb conferring resistance to ToLCNDV-ES, particularly resistance to resistance-destroying strains, and wherein the introgression segment contains a gene encoding a protein of SEQ ID NO: 8, or a gene encoding a protein having at least 96% sequence identity with SEQ ID NO: 8, and wherein said protein preferably contains at least one, preferably at least two, three, or all four of the following amino acids:

[0272] - Nine serines (S) at equivalent positions in amino acids 10 to 18 of SEQ ID NO: 8, or in a sequence having at least 96% sequence identity with SEQ ID NO: 8.

[0273] - Methionine (M) at amino acid number 484 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0274] - Lysine (K) at amino acid number 629 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0275] - Histidine (H) or aspartic acid (D) at amino acid number 679 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0276] In one respect, QTL5rb (or introgression fragments containing it or potential causal genes of QTL5rb) may be obtained from (or may be obtained from, derived from or may be derived from, or exist in) seeds whose representative samples have been deposited with accession number NCIMB 44139.

[0277] The aforementioned cultivated melon plants and plant cells contain an introgression fragment comprising the sequence of nucleotide 26034368 originating from chromosome 5 and nucleotide 26027838 terminating from chromosome 5 of the ToLCNDV-ES resistance donor melon plant (this region corresponds to the region of SEQ ID NO: 11 or a variant thereof found in reference melonomics.net, V4.0), and optionally wherein the introgression fragment is obtained from seeds whose representative samples have been deposited with accession number NCIMB44139.

[0278] In one aspect, the aforementioned cultivated melon plants and plant cells comprise SEQ ID NO: 11 or a variant thereof, said variant having at least 95%, 96%, 97%, 98%, or 99% sequence identity with SEQ ID NO: 11, and / or said variant encodes a protein preferably containing at least one of the following amino acids, preferably at least two, three, or all four amino acids:

[0279] - Nine serines (S) at equivalent positions in amino acids 10 to 18 of SEQ ID NO: 8, or in a sequence having at least 96% sequence identity with SEQ ID NO: 8.

[0280] - Methionine (M) at amino acid number 484 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0281] - Lysine (K) at amino acid number 629 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0282] - Histidine (H) or aspartic acid (D) at amino acid number 679 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8.

[0283] In another aspect, the aforementioned cultivated melon plants and plant cells are provided, wherein the introgression fragment comprises one or more SNP markers selected from SNP_02, SNP_03, SNP_04, SNP_05, and SNP_06, particularly one or more SNP markers selected from SNP_02, SNP_03, SNP_04, and SNP_05, particularly donor SNP nucleotides of at least SNP_03 and / or SNP_04 or at least SNP_03, SNP_04, and SNP_05. Thus, SNP haplotypes, for example, are... Figure 3 As shown: GGTAT (for SNP_02 to SNP_06) or, for example, GGTA (for SNP_02 to SNP_05) or GT (for SNP_03 and SNP_04) or G and / or T (for SNP_03 and / or SNP_04).

[0284] In another aspect, the aforementioned cultivated melon plants and plant cells are provided, wherein the infiltrated fragment comprises a donor SNP nucleotide selected from one or more SNP markers, such as CGTATC of SNPs 01 to 07, including SNPs 01, 02, 03, 04, 05, 06, and 07, for example: CGTATC of SNPs 01 to 07, see, for example Figure 3 .

[0285] In one aspect, the cultivated melon plant or plant cell contains a homozygous introgression fragment and contains, for example, the genotype GG-GG-TT-AA-TT (for SNP_02 to SNP_06) or, for example, GG-GG-TT-AA (for SNP_02 to SNP_05) or GG-TT (for SNP_03 and SNP_04) or GG and / or TT (for SNP_03 and / or SNP_04).

[0286] In another aspect, a melon plant or plant cell is provided, wherein the infiltrated fragment further comprises guanine (G) at nucleotide 81 of SEQ ID NO: 3 or at an equivalent nucleotide of a sequence having at least 95%, 96%, 97%, 98%, or 99% sequence identity with SEQ ID NO: 3, and / or adenine (A) at nucleotide 101 of SEQ ID NO: 4 or at an equivalent nucleotide of a sequence having at least 95%, 96%, 97%, 98%, or 99% sequence identity with SEQ ID NO: 4, or thymine (T) at nucleotide 101 of the complementary strand of SEQ ID NO: 4 or at nucleotide 101 of the complementary strand of SEQ ID NO: 4.

[0287] A seed is also provided from which a cultivated melon plant as described in this article can be grown.

[0288] Cultivated melon seeds contain plant cells containing QTL5rb or a potential causal gene for QTL5rb, as described in this article.

[0289] In addition, cultivated melon fruits containing plant cells as described herein, said plant cells containing QTL5rb or a potential causal gene of QTL5rb, are also provided. Fruits produced on said melon plants and containing at least one chromosome 5 containing a potential causal gene of QTL5rb are also provided.

[0290] Also provided is propagation material of cultivated melon plants containing plant cells as described herein, said plant cells containing QTL5rb or a potential causal gene QTL5rb.

[0291] A method for generating ToLCNDV-ES resistant melon plants containing QTL5rb or a potential causal gene of QTL5rb is also provided, the method comprising the following steps:

[0292] a) Select ToLCNDV-ES resistant donor plants, especially those resistant to ToLCNDV-ES strains that destroy resistance;

[0293] b) Hybridize the donor plant selected in step a) with plants susceptible to ToLCNDV-ES, especially those susceptible to ToLCNDV-ES strains that destroy resistance.

[0294] c) Obtain F1 seeds from the hybridized plants of step b), or seeds of further self-pollinating generations such as F2, F3, or further self-pollinating generations, or seeds of backcross generations obtained by backcrossing ToLCNDV-ES resistant plants with ToLCNDV-ES sensitive plants, and optionally...

[0295] d) Verify whether plants grown from the seeds obtained in step c) are resistant to ToLCNDV-ES (especially to strains that destroy resistance) and / or contain:

[0296] - One or more of the following donor SNP haplotypes: SNP_01 to SNP_07, or SNP_02 to SNP_06, or SNP_03 to SNP_05, or SNP_03 to SNP_04; and / or

[0297] - Guanine (G) at nucleotide 81 of SEQ ID NO: 3 or at an equivalent nucleotide of a sequence having at least 95%, 96%, 97%, 98%, or 99% sequence identity with SEQ ID NO: 3, and / or adenine (A) at nucleotide 101 of SEQ ID NO: 4 or at an equivalent nucleotide of a sequence having at least 95%, 96%, 97%, 98%, or 99% sequence identity with SEQ ID NO: 4, or thymine (T) at nucleotide 101 of the complementary strand of SEQ ID NO: 4 or at nucleotide 101 of the complementary strand of SEQ ID NO: 4; and / or

[0298] - A gene encoding the protein of SEQ ID NO: 8, or a gene encoding a protein having at least 96% sequence identity with SEQ ID NO: 8, wherein said protein preferably comprises at least one of the following, preferably at least two, three, or all four:

[0299] - Nine serines (S) at equivalent positions in amino acids 10 to 18 of SEQ ID NO: 8, or in a sequence having at least 96% sequence identity with SEQ ID NO: 8.

[0300] - Methionine (M) at amino acid number 484 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0301] - Lysine (K) at amino acid number 629 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0302] - Histidine (H) or aspartic acid (D) at amino acid number 679 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8; and / or

[0303] - The nucleotide sequence of SEQ ID NO: 11 or having at least 95%, 96%, 97%, 98% or 99% sequence identity with SEQ ID NO: 11, preferably encoding a protein as described above.

[0304] The verification step in step d) can optionally be performed additionally or alternatively in step a), i.e., when the donor plant is selected. Therefore, the donor plant can be selected, for example, based on SNP markers and / or based on the gene sequence or nucleotide sequence of step d).

[0305] In one aspect, the introgression fragment containing QTL5rb (or the potential causal gene of QTL5rb) may be obtained (or obtained from, may be derived from, originate from, etc.) from seeds preserved with NCIMB 44139 or their progeny, said progeny retaining QTL5rb (or the potential causal gene of QTL5rb).

[0306] A method for producing melon seeds is also provided, the method comprising the following steps:

[0307] a) Develop a melon plant containing at least one chromosome 5, wherein the at least one chromosome 5 has an introgression segment from a ToLCNDV-ES resistance donor plant, wherein the introgression segment contains a QTL5rb (or a potential causal gene for QTL5rb) conferring ToLCNDV-ES resistance and contains:

[0308] - One or more of the following donor SNP haplotypes: SNP_01 to SNP_07, or SNP_02 to SNP_06, or SNP_03 to SNP_05, or SNP_03 to SNP_04; and / or

[0309] - Guanine (G) at nucleotide 81 of SEQ ID NO: 3 or at an equivalent nucleotide of a sequence having at least 95%, 96%, 97%, 98%, or 99% sequence identity with SEQ ID NO: 3, and / or adenine (A) at nucleotide 101 of SEQ ID NO: 4 or at an equivalent nucleotide of a sequence having at least 95%, 96%, 97%, 98%, or 99% sequence identity with SEQ ID NO: 4, or thymine (T) at nucleotide 101 of the complementary strand of SEQ ID NO: 4 or at nucleotide 101 of the complementary strand of SEQ ID NO: 4; and / or

[0310] - A gene encoding the protein of SEQ ID NO: 8, or a gene encoding a protein having at least 96% sequence identity with SEQ ID NO: 8, wherein said protein preferably contains at least one of the following amino acids, preferably at least two, three, or all four amino acids:

[0311] - Nine serines (S) at equivalent positions in amino acids 10 to 18 of SEQ ID NO: 8, or in a sequence having at least 96% sequence identity with SEQ ID NO: 8.

[0312] - Methionine (M) at amino acid number 484 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0313] - Lysine (K) at amino acid number 629 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0314] - Histidine (H) or aspartic acid (D) at amino acid number 679 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8; and / or

[0315] - The nucleotide sequence of SEQ ID NO: 11 or having at least 95%, 96%, 97%, 98%, or 99% sequence identity with SEQ ID NO: 11, preferably encoding a protein as described above.

[0316] b) Harvest the fruits of the melon plants grown in step a), and optionally...

[0317] c) Collect seeds from the fruit obtained in step b).

[0318] In one aspect, the introgression fragment containing QTL5rb (or a potential causal gene for QTL5rb) may be obtained from (or may be derived from, or may be present in) a seed deposited with NCIMB 44139 or its progeny, which retains QTL5rb.

[0319] In another aspect, a method for producing hybrid melon seeds is provided, the method comprising the following steps:

[0320] a) To produce a first-inbred melon plant containing at least one chromosome 5 with an introgression segment including QTL5rb as described herein.

[0321] Hybridization with second-inbred melon plants, with or without an introgression fragment on chromosome 5 containing QTL5rb as described herein, and

[0322] b) Select the seeds obtained from the hybridization in step a).

[0323] In another aspect, a method for producing melon fruit is provided, the method comprising the following steps:

[0324] a) To grow a melon plant containing at least one chromosome 5 with an introgression segment as described herein, and

[0325] b) Harvest the fruit produced by the plants grown in step a).

[0326] Resistance to ToLCNDV-ES conferred by the introgression fragment or QTL5rb is expressed in a dominant manner, and therefore can be observed when there is only one chromosome 5 in the plant containing the introgression fragment or QTL5rb (or the potential causal gene of QTL5rb).

[0327] Therefore, other specific embodiments of the invention relate to melon plant cells or melon plants in which an introgression fragment from chromosome 5 of a ToLCNDV-ES resistance donor plant containing sequences between SNP_01 and SNP_07, between SNP_07 and SNP_06, between SNP_02 and SNP_05, between SNP_03 and SNP_06, between SNP_03 and SNP_05, between SNP_02 and SNP_04, between SNP_03 and SNP_04, between SNP_03 and SNP_05, or preferably between SNP_03 and SNP_04, exists in a heterozygous state. Thus, it is sufficient for at least one chromosome 5 in the plant cell or plant to contain the introgression fragment just described. However, it should be understood that, due to the dominance of ToLCNDV-ES resistance conferred by the introgression fragment just described, chromosome 5 with respect to the introgression fragment just described can also exist in a homozygous state without reducing the degree of resistance. Therefore, the present invention includes plant cells according to the invention or plants according to the invention, which contain the introgression fragments just described in a heterozygous or homozygous state.

[0328] Table 1 illustrates the SNP genotypes and haplotypes of plant or cell SNPs containing homozygous (on both chromosomes 5) or heterozygous (on one chromosome 5) ToLCNDV-ES resistance donor SNPs. In the table, SNP genotype or haplotype refers to the forward strand relative to the reference genome.

[0329] Table 1

[0330] SNPs and nucleotide positions (nt) in the sequence SNP genotypes in melon plants containing homozygous donor fragments SNP haplotypes in melon plants containing heterozygous donor fragments SNP genotypes in melon plants containing heterozygous donor fragments, where X can be any nucleotide, such as a nucleotide reverting to the parent. SNP_01 (nucleotide 81* of SEQ ID NO 1) CC C CX SNP_02 (nucleotide 242* of SEQ ID NO 2) CC (or GG when referring to the complementary chain) C (or G when referring to the complementary chain) CX (or GX when referring to the complementary chain) SNP_03 (nucleotide 81* of SEQ ID NO 3) GG G GX SNP_04 (nucleotide 101* of SEQ ID NO 4) AA (or TT when referring to the complementary chain) A (or T when referring to the complementary chain) AX (or TX when referring to the complementary chain) SNP_05 (nucleotide 101* of SEQ ID NO 5) TT (or AA when referring to the complementary chain) T (or A when referring to the complementary chain) TX (or AX when referring to the complementary chain) SNP_06 (nucleotide 101* of SEQ ID NO 6) AA (or TT when referring to the complementary chain) A (or T when referring to the complementary chain) AX (or TX when referring to the complementary chain) SNP_07 (nucleotide 81* of SEQ ID NO 7) CC C CX

[0331] *Or nucleotides (nt) at equivalent positions in sequences that have at least 95%, 96%, 97%, 98% or 99% sequence identity with a given sequence (or complementary sequence).

[0332] In one embodiment, a cultivated melon plant (melon species) is provided, the cultivated melon plant containing recombinant chromosome 5, whereby the recombinant chromosome 5 contains an introgression fragment that confers ToLCNDV-ES resistance to the melon plant when present in homozygous or heterozygous form, and wherein the introgression fragment originates from a wild-type donor of the melon species. In one aspect, the introgression fragment comprises one or more of SNP_01, SNP_02, SNP_03, SNP_04, SNP_05, SNP_06, and / or SNP_07, optionally one or more of SNP_02, SNP_03, SNP_04, SNP_05, and / or SNP_06 donor haplotypes. In one embodiment, the introgression fragment comprises at least SNP_03 and / or SNP_04, or at least SNP_03, SNP_04, and SNP_05 donor SNP haplotypes. The donor SNP haplotype can be present in a homozygous form (if the introgression fragment is homozygous) or a heterozygous form (if the introgression fragment is heterozygous). Therefore, for example, a plant, plant cell, or plant part can contain the GG or GX genotype of SNP_03 at nucleotide 81 of SEQ ID NO: 3 or at an equivalent nucleotide in a sequence having at least 95% sequence identity with SEQ ID NO: 3. Therefore, a plant, plant cell, or plant part can contain either a homozygous or heterozygous form of SEQ ID NO: 3 or a sequence having at least 95%, 97%, 98%, or 99% sequence identity with SEQ ID NO: 3.

[0333] In one embodiment, a cultivated melon plant or cells of such plants are provided containing an introgression fragment from a wild-type donor on chromosome 5, said introgression fragment conferring ToLCNDV-ES resistance (particularly resistance to strains that destroy resistance), wherein the introgression fragment is located between SNP_02 and SNP_06 (or between SEQ ID NO: 2 and SEQ ID NO: 6, or between sequences having at least 95%, or at least 97%, or 98%, or 99% sequence identity with SEQ ID NO: 2 or SEQ ID NO: 6); or wherein the introgression fragment is located between SNP_02 and SNP_05 (or between SEQ ID NO: 2 and SEQ ID NO: 5, or between sequences having at least 95%, or at least 97%, or 98%, or 99% sequence identity with SEQ ID NO: 2 or SEQ ID NO: 5); or wherein the introgression fragment is located between SNP_03 and SNP_04 (or between SEQ ID NO: 3 and SEQ ID NO: 4, or between SEQ ID NO: 6 and SEQ ID NO: 6). The infiltrating fragment is located between SNP_03 and SNP_05 (or between SEQ ID NO: 3 and SEQ ID NO: 5, or between sequences having at least 95%, or at least 97%, or 98%, or 99% sequence identity with SEQ ID NO: 3 or SEQ ID NO: 5).

[0334] In one aspect, cultivated melon plants or cells of such plants contain an introgression segment from a wild-type donor on chromosome 5, said introgression segment conferring resistance to ToLCNDV-ES (particularly resistance to strains that destroy resistance), wherein the introgression segment is located between SNP_02 and SNP_06 (or between SEQ ID NO: 2 and SEQ ID NO: 6, or between sequences having at least 95%, or at least 97%, or 98%, or 99% sequence identity with SEQ ID NO: 2 or SEQ ID NO: 6), or the segment is located between SNP_02 and SNP_05 (or between SEQ ID NO: 2 and SEQ ID NO: 5, or between sequences having at least 95%, or at least 97%, or 98%, or 99% sequence identity with SEQ ID NO: 2 or SEQ ID NO: 5), or thereby the introgression segment is located between SNP_03 and SNP_04 (or between SEQ ID NO: 3 and SEQ ID NO: 4, or between SEQ ID NO: 3 or SEQ ID NO: 4). 4. Between sequences having at least 95%, or at least 97%, or 98%, or 99% sequence identity, or whereby the infiltrated fragment is located between SNP_03 and SNP_05 or between sequences having at least 95%, or at least 97%, or 98%, or 99% sequence identity with SEQ ID NO: 3 or SEQ ID NO: 5, and / or whereby the infiltrated fragment contains a gene encoding the protein of SEQ ID NO: 8, or a gene encoding a protein having at least 96% sequence identity with SEQ ID NO: 8, and wherein said protein preferably comprises at least one of the following, preferably at least two, three, or all four:

[0335] - Nine serines (S) at equivalent positions in amino acids 10 to 18 of SEQ ID NO: 8, or in a sequence having at least 96% sequence identity with SEQ ID NO: 8.

[0336] - Methionine (M) at amino acid number 484 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0337] - Lysine (K) at amino acid number 629 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0338] - Histidine (H) or aspartic acid (D) at amino acid number 679 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0339] And / or

[0340] - The nucleotide sequence of SEQ ID NO: 11 or having at least 95%, 96%, 97%, 98% or 99% sequence identity with SEQ ID NO: 11, preferably encoding a protein as described above.

[0341] In another aspect, the cultivated melon plants or the cells of these plants contain an introgression segment from a wild-type donor on chromosome 5, which confers resistance to ToLCNDV-ES (especially resistance to strains that destroy resistance), wherein the introgression segment contains a gene encoding the protein of SEQ ID NO: 8, or a gene encoding a protein having at least 96% sequence identity with SEQ ID NO: 8, and wherein said protein preferably comprises at least one, preferably at least two, three, or all four of the following:

[0342] - Nine serines (S) at equivalent positions in amino acids 10 to 18 of SEQ ID NO: 8, or in a sequence having at least 96% sequence identity with SEQ ID NO: 8.

[0343] - Methionine (M) at amino acid number 484 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0344] - Lysine (K) at amino acid number 629 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0345] - Histidine (H) or aspartic acid (D) at amino acid number 679 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0346] And / or

[0347] - The nucleotide sequence of SEQ ID NO: 11 or having at least 95%, 96%, 97%, 98% or 99% sequence identity with SEQ ID NO: 11, preferably encoding a protein as described above.

[0348] In another aspect, the cultivated melon plants or the cells of these plants contain an endogenous gene on chromosome 5 encoding a TIR-NBS-LRR protein that confers resistance to ToLCNDV-ES (especially resistance to strains that destroy resistance), whereby this gene encodes the protein of SEQ ID NO: 8, or a protein having at least 96% sequence identity with SEQ ID NO: 8, and wherein said protein preferably comprises at least one, preferably at least two, three, or all four of the following:

[0349] - Nine serines (S) at equivalent positions in amino acids 10 to 18 of SEQ ID NO: 8, or in a sequence having at least 96% sequence identity with SEQ ID NO: 8.

[0350] - Methionine (M) at amino acid number 484 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0351] - Lysine (K) at amino acid number 629 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0352] - Histidine (H) or aspartic acid (D) at amino acid number 679 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0353] And / or

[0354] -The gene contains the nucleotide sequence of SEQ ID NO: 11 or has at least 95%, 96%, 97%, 98% or 99% sequence identity with SEQ ID NO: 11, and preferably encodes a protein as described above.

[0355] In one aspect, cultivated melon plants, plant cells, or plant parts containing the modified endogenous TIR-NBS-LRR gene described above and elsewhere herein on chromosome 5 are provided. The advantage of modifying the endogenous gene in a cultivated melon line rather than introgressing a gene from a donor into the cultivated melon is that the genetic background of the melon plant is the cultivar background, and no linkage drag is introduced into the cultivar background, nor is lengthy backcrossing required. This eliminates the need for numerous generations of backcrossing with superior parent lines and the removal of unwanted linkage drag.

[0356] In one respect, the endogenous gene has been modified by targeted mutagenesis and / or mutated by random mutagenesis so that the gene does not encode a susceptible TIR-NBS-LRR protein (such as the susceptible protein of SEQ ID NO: 10, or a susceptible protein having at least 98%, 98.5%, 99%, 99.5% or 99.8% sequence identity with SEQ ID NO: 10).

[0357] Therefore, in one aspect, a cultivated melon plant, plant cells, and plant parts are provided, wherein an endogenous gene has been modified by targeted mutagenesis and / or mutated by random mutagenesis to confer QTL5rb resistance, thereby encoding the resistance-conferring protein of SEQ ID NO: 8, or a protein having at least 96%, 97%, 98%, or 99% sequence identity with the protein of SEQ ID NO: 8, and said protein preferably contains one or more of the following four amino acid variations compared to the susceptibility protein of SEQ ID NO: 10:

[0358] - "Change 1": At the beginning of the protein, two S (serine) residues are inserted after amino acid E9 (glutamic acid 9); that is, the protein contains 9 serine residues (S) after E9, and amino acids 10 to 18 are serine residues in proteins that have at least 96%, 97%, 98%, or 99% sequence identity with the protein of SEQ ID NO: 8 (while SEQ ID NO: 10 contains only 7 serine residues, i.e., amino acids 10 to 16); see also Figure 2 Change number 1;

[0359] - "Change No. 2": At the equivalent position in a protein having at least 96%, 97%, 98%, or 99% sequence identity with the protein of SEQ ID NO: 8, amino acid V483 (valine 483 of SEQ ID NO: 10) is changed to M (methionine); in SEQ ID NO: 8, the equivalent position is M484; see also Figure 2 Change number 2;

[0360] - "Change No. 3": Amino acid E628 (glutamic acid 628 of SEQ ID NO: 10) is changed to K (lysine) at the equivalent position in proteins having at least 96%, 97%, 98%, or 99% sequence identity with the protein of SEQ ID NO: 8; in SEQ ID NO: 8, the equivalent position is K629; see also Figure 2 Change number 3;

[0361] - "Change No. 4": At the equivalent position in a protein having at least 96%, 97%, 98%, or 99% sequence identity with the protein of SEQ ID NO: 8, amino acid Y678 (tyrosine 678 of SEQ ID NO: 10) is changed to H (histidine); in SEQ ID NO: 8, the equivalent position is H679, see [link to relevant documentation]. Figure 2 Change 4; or at the equivalent position in a protein having at least 96%, 97%, 98%, or 99% sequence identity with the protein of SEQ ID NO: 8, amino acid Y678 (tyrosine 678 of SEQ ID NO: 10) is changed to D (aspartic acid); in SEQ ID NO: 8, the equivalent position is D679, see [link to SEQ ID NO: 8]. Figure 1 or Figure 2 Change number 4.

[0362] In one aspect, one or more of the variations 1 through 4 are introduced through targeted mutagenesis or targeted gene editing (such as CRISPR-based gene editing) and / or through random mutagenesis (such as radiation treatment or chemical mutagenesis). For example, Figure 4 Nucleotide changes in the genome sequence are shown. These changes can be generated by random mutagenesis or targeted mutagenesis to insert one or more amino acid codons (e.g., change 1) or to replace codons with different codons (e.g., change 2 to change 4).

[0363] Therefore, modified or mutated cultivated melon plants, plant parts, cells, fruits, tissues, propagation materials, or seeds do not contain the introgression fragment of QTL5rb, but contain a modified endogenous gene at a locus on chromosome 5 that encodes a "resistance protein" and confers resistance to ToLCNDV-ES strains, especially resistance-destroying strains.

[0364] Plants and plant parts containing plant cells according to the invention (e.g., containing introgression fragments or containing modified or mutated endogenous genes) are another embodiment of the invention. Seeds capable of growing such plants are also provided herein.

[0365] Therefore, this article also covers, for example, the introgression of QTL5rb (or a potential causal gene for QTL5rb) into fruits, seeds, plants, plant cells, plant tissues, and vegetative propagation, for the purpose of modifying endogenous TIR-NBS-LRR genes (encoding susceptibility proteins) through targeted or random mutagenesis. This covers methods for growing such plants, methods for selecting such plants or plant parts, and methods for producing hybrid or inbred lines containing one or two copies of the modified or mutated endogenous TIR-NBS-LRR gene.

[0366] The melon plants described herein (e.g., containing introgression fragments or modified or mutated endogenous genes) can be inbred lines, open-pollinated varieties (OP), or F1 hybrids. In one aspect, F1 hybrids contain heterozygous introgression fragments or modified endogenous genes, for example, produced by crossing two inbred parent lines (one of which has an introgression fragment or modified endogenous gene (preferably homozygous, but not necessarily)) and collecting F1 hybrid seeds from said cross. F1 hybrids may also contain homozygous introgression fragments or modified endogenous genes, i.e., produced by crossing two inbred parent lines, each containing either homozygous or heterozygous introgression fragments or modified endogenous genes.

[0367] The melon plant described herein can be of any type. Preferably, the melon plant has good agronomic characteristics and good fruit quality characteristics, such as large average fruit size (at least 500g, 600g, 700g, 800g, 900g, 1000g or larger), high average Brix content of the fruit (e.g., average refractometer % total soluble solids of at least 10%, 12%, 14%, 16%, 18% or higher), many fruits per plant, and firm flesh.

[0368] Other resistances can also be introduced into the melon plants described herein, such as resistance to one or more of the following diseases: bacterial wilt, root rot, crown wilt, melon rust, powdery mildew, Verticillium wilt, sulfur burn, scab, watermelon mosaic, downy mildew, Fusarium oxysporum race 0, Fusarium oxysporum race 1, Fusarium oxysporum race 2, Fusarium oxysporum race 1.2, Fusarium wilt R2, root-knot nematode, anthracnose, cucumber mosaic and pumpkin mosaic, and / or resistance to one or more of the following pests: aphid resistance, pickle beetle, false ground beetle, cucumber band beetle, mites, western spotted cucumber beetle, melon leafhopper, melon worm, western striped cucumber beetle, or melon leafminer. Other resistance genes against pathogenic viruses, fungi, bacteria, or pests can also be introduced.

[0369] One specific aspect of the invention relates to plants or plant cells containing an introgression fragment, which may be obtained from seeds or their progeny preserved under NCIMB 44139. The preserved seeds are those of wild-type donors containing a homozygous form of QTL5rb, as used in the examples, wherein the donor nucleotides are present in homozygous forms of SNP_01 to SNP_07. The causal gene for ToLCNDV-ES resistance is located between SNP_03 and SNP_04, meaning that the size of the donor gene introgression can be small by selecting recombinants with a small introgression fragment, such as a causal gene encoding only the protein of SEQ ID NO: 8.

[0370] Whether a plant contains ToLCNDV-ES resistance from preserved seeds can be determined by various methods, such as sequencing.

[0371] Melon plants containing the melon plant cells according to the present invention, and plant parts (such as leaves, stems, roots, fruits, pollen, flowers, etc.) are also embodiments of the present invention. Similarly, this document covers seeds that can grow such plants, as well as parts of such seeds (e.g., seed cells or tissues, such as seed coat, embryo, etc.).

[0372] Another aspect of the invention relates to melon seeds or plants comprising an introgression fragment from chromosome 5 of a ToLCNDV-ES resistant donor plant, wherein the introgression fragment comprises a sequence from the ToLCNDV-ES resistant donor melon plant between SNP_01 and SNP_07, between SNP_02 and SNP_06, between SNP_02 and SNP_05, between SNP_02 and SNP_04, or between SNP_03 and SNP_04. In a preferred embodiment of the invention, the seed contains an introgression fragment from a ToLCNDV-ES resistant donor plant on chromosome 5, wherein the introgression fragment contains a sequence from the ToLCNDV-ES resistant melon plant between SNP_02 and SNP_06, more preferably between SNP_02 and SNP_05 or between SNP_03 and SNP_05, even more preferably between SNP_02 and SNP_04, and most preferably between SNP_03 and SNP_04.

[0373] Therefore, it encompasses a melon plant, seed, or plant part containing an introgression fragment from a ToLCNDV-ES resistance donor plant on chromosome 5, wherein the introgression fragment contains a sequence from a ToLCNDV-ES resistance donor melon plant, the sequence containing QTL5rb (or at least a potential causal gene of QTL5rb) and containing one or more of SNP_01 to SNP_07, optionally one or more of SNP_02 to SNP_06, such as one or more of SNP_02 to SNP_05 or SNP_03 to SNP_05, or one or more of SNP_02 to SNP_04, or donor SNP nucleotides of SNP_03 and / or SNP_04.

[0374] Since the causal gene of QTL5rb is located between SNP_03 and SNP_04 on chromosome 5, one aspect of this document is that, for example, when this document refers to an introgression fragment or QTL5rb between SNP_03 and SNP_04, this covers a fragment or region of a plant cell, seed, fruit, propagation material, tissue, or plant that contains a gene encoding the protein of SEQ ID NO: 8 or a gene encoding a protein having at least 96% sequence identity with SEQ ID NO: 8, and said protein preferably comprises at least one, preferably at least two, three, or all four of the following:

[0375] - Nine serines (S) at equivalent positions in amino acids 10 to 18 of SEQ ID NO: 8, or in a sequence having at least 96% sequence identity with SEQ ID NO: 8.

[0376] - Methionine (M) at amino acid number 484 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0377] - Lysine (K) at amino acid number 629 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0378] - Histidine (H) or aspartic acid (D) at amino acid number 679 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0379] And / or the fragment contains the nucleotide sequence of SEQ ID NO: 11 or has at least 95%, 96%, 97%, 98% or 99% sequence identity with SEQ ID NO: 11, and preferably encodes a protein as described above.

[0380] Instead of detecting one or more donor SNP nucleotides from SNP_01 to SNP_07, preferably SNP_02 to SNP_06, this implementation also covers the detection and / or selection and / or verification of plant cells, seeds, fruits, propagation materials, tissues or plants based on the presence of potential causal genes in QTL5rb.

[0381] Another embodiment of the invention relates to melon seeds that are available from or derived from plants according to the invention, or seeds containing plant cells according to the invention.

[0382] Another aspect of the invention relates to melon fruit, plant, cell, seed, or plant tissue comprising cells containing, for example, an introgression fragment from a ToLCNDV-ES resistant donor plant on chromosome 5, wherein the introgression fragment comprises a sequence from a ToLCNDV-ES resistant donor melon plant between SNP_01 and SNP_07, between SNP_02 and SNP_06, between SNP_02 and SNP_05, between SNP_03 and SNP_05, between SNP_02 and SNP_04, or between SNP_03 and SNP_04.

[0383] In another embodiment, the seed, plant, cell, fruit, or plant tissue contains an introgression fragment from a ToLCNDV-ES resistance donor plant on chromosome 5, wherein the introgression fragment contains a sequence from the ToLCNDV-ES resistance donor melon plant between SNP_01 and SNP_07, more preferably between SNP_02 and SNP_06, even more preferably between SNP_02 and SNP_05 or between SNP_03 and SNP_05, further preferably between SNP_02 and SNP_04, and most preferably between SNP_03 and SNP_04.

[0384] In another embodiment, the plant, cell, seed, fruit, or plant tissue contains an introgression fragment on chromosome 5 from a ToLCNDV-ES resistance donor plant, wherein the introgression fragment contains a sequence from a ToLCNDV-ES resistance donor melon plant, the sequence containing a QTL5rb (or at least a potential causal gene for QTL5rb) and containing one or more of SNP_01 to SNP_07, optionally one or more of SNP_02 to SNP_06, such as one or more of SNP_02 to SNP_05 or one or more of SNP_03 to SNP_05, or one or more of SNP_02 to SNP_04, or donor SNP nucleotides of SNP_03 and / or SNP_04.

[0385] Another embodiment of the invention relates to melon fruit that is available from or obtained from a plant according to the invention, or fruit containing plant cells according to the invention.

[0386] Preferably, the melon fruits provided in one aspect of this document are characterized in that they contain an introgression fragment from a ToLCNDV-ES resistant donor plant on chromosome 5, the introgression fragment containing a sequence of the donor plant in a heterozygous or homozygous state between SNP_01 and SNP_07, between SNP_02 and SNP_06, between SNP_02 and SNP_05, between SNP_02 and SNP_04, between SNP_03 and SNP_06, between SNP_03 and SNP_05, or preferably between SNP_03 and SNP_04.

[0387] Therefore, the preferred and further embodiments described herein for melon plant cells or melon plants according to the present invention also apply to the preferred and further embodiments of melon fruits representing melon plants according to the present invention.

[0388] Another aspect provided in this paper relates to melon plant propagation material containing an introgression fragment from a ToLCNDV-ES resistance donor plant on chromosome 5, wherein the introgression fragment contains a sequence from the ToLCNDV-ES resistance donor melon plant between SNP_01 and SNP_07, between SNP_02 and SNP_05, between SNP_02 and SNP_04, between SNP_03 and SNP_06, between SNP_03 and SNP_05, or between SNP_03 and SNP_04. Therefore, melon plant propagation material containing an introgression fragment from a ToLCNDV-ES resistant donor plant on chromosome 5, wherein the introgression fragment contains a sequence from a ToLCNDV-ES resistant donor melon plant, the sequence containing QTL5rb (or at least a potential causal gene of QTL5rb) and containing one or more of SNP_01 to SNP_07, optionally one or more of SNP_02 to SNP_06, such as one or more of SNP_02 to SNP_05 or one or more of SNP_03 to SNP_05, or one or more of SNP_02 to SNP_04, or donor SNP nucleotides of SNP_03 and / or SNP_04.

[0389] Another embodiment of the invention relates to melon plant propagation material that is available from or derived from plants according to the invention, or melon plant propagation material containing plant cells according to the invention.

[0390] Therefore, the preferred and further embodiments of the plant cells or plant descriptions according to the present invention are also applicable to the preferred and further embodiments of the propagation material representing the melon plant according to the present invention.

[0391] The term "propagation material" includes those components of a plant suitable for producing offspring via vegetative (asexual) or reproductive (sexual) pathways. Suitable for vegetative propagation include, for example, cuttings, in vitro tissues, cells, protoplasts, embryos or callus cultures, micropropagation methods, rhizomes, or tubers. Other propagation materials include, for example, fruits, seeds, seedlings, and those that are homozygous or heterozygous for introgression fragments on chromosome 5 or for modified endogenous genes (e.g., modified by editing or random mutagenesis) conferring resistance to ToLCNDV-ES. In one aspect, propagation material takes the form of cuttings, which are propagated by grafting onto another rootstock or in vitro tissue culture material, especially embryo cultures. Propagation material in the form of in vitro tissue culture material, particularly in vitro embryo cultures, is particularly preferred.

[0392] In one aspect, non-reproductive plant cells containing recombinant chromosome 5 or modified endogenous genes as described herein are provided.

[0393] Another embodiment of the invention relates to a method for producing ToLCNDV-ES resistant melon plants (especially resistant to strains that destroy resistance), the method comprising the following steps

[0394] a) Select ToLCNDV-ES resistance donor plants

[0395] b) Hybridize the donor plant selected in step a) with a regenerating plant sensitive to ToLCNDV-ES.

[0396] c) Obtain seeds from the hybridized plants of step b), and optionally...

[0397] d) Verify whether the plants grown from the seeds obtained in step c) are resistant to TOLCKEV-ES and / or contain one or more SNPs from the donor plant selected from the group consisting of SNP_01, SNP_02, SNP_03, SNP_04, SNP_05, SNP_06 and SNP_07, especially selected from SNP_02 to SNP_06;

[0398] And / or a gene encoding the protein of SEQ ID NO: 8, or a gene encoding a protein having at least 96% sequence identity with SEQ ID NO: 8, wherein said protein preferably contains at least one of the following amino acids, preferably at least two, three, or all four amino acids:

[0399] - Nine serines (S) at equivalent positions in amino acids 10 to 18 of SEQ ID NO: 8, or in a sequence having at least 96% sequence identity with SEQ ID NO: 8.

[0400] - Methionine (M) at amino acid number 484 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0401] - Lysine (K) at amino acid number 629 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0402] - Histidine (H) or aspartic acid (D) at amino acid number 679 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0403] And / or the nucleotide sequence of SEQ ID NO: 11 or having at least 95%, 96%, 97%, 98% or 99% sequence identity with SEQ ID NO: 11, preferably a sequence encoding the above-mentioned protein.

[0404] In step a) of the method for producing ToLCNDV-ES resistant melon plants according to the invention, the ToLCNDV-ES resistant donor plants can be selected by infecting the melon plants with ToLCNDV-ES, particularly with a strain that disrupts resistance, and determining the symptomatic level of the ToLCNDV-ES-infected melon plants as described elsewhere herein. The same applies to step c) of the method for producing ToLCNDV-ES resistant melon plants according to the invention, if the plants are either ToLCNDV-ES susceptible or resistant.

[0405] In a preferred embodiment of the method for producing ToLCNDV-ES resistant melon plants, the ToLCNDV-ES resistant donor plant in step a) contains a ToLCNDV-ES resistance-conferring segment on chromosome 5, which comprises a sequence of the donor plant between SNP_01 and SNP_07, between SNP_02 and SNP_06, between SNP_02 and SNP_05, between SNP_02 and SNP_04, between SNP_03 and SNP_06, between SNP_03 and SNP_05, or between SNP_03 and SNP_04. Most preferably, the ToLCNDV-ES resistance-conferring segment on chromosome 5 comprises a sequence between SNP_03 and SNP_04. In a particularly preferred embodiment of the invention, the method for producing ToLCNDV-ES resistant melon plants according to the invention, in step a), the ToLCNDV-ES resistant donor plant contains a segment on chromosome 5 conferring ToLCNDV-ES resistance, the segment comprising sequences found in (available from) seeds preserved with NCIMB 44139 between SNP_01 and SNP_07, between SNP_02 and SNP_06, between SNP_02 and SNP_05, between SNP_02 and SNP_04, between SNP_03 and SNP_06, between SNP_03 and SNP_05, or between SNP_03 and SNP_04.

[0406] In a preferred embodiment of the invention, the method for producing ToLCNDV-ES resistant melon plants according to the invention is used to produce plants comprising a homozygous or heterozygous QTL5rb. The preferred and further embodiments described herein with respect to plants according to the invention are therefore applicable to methods for producing ToLCNDV-ES resistant melon plants comprising QTL5rb.

[0407] The plant obtained by the method for producing ToLCNDV-ES resistant melon plants according to the present invention is also an embodiment of the present invention.

[0408] Another embodiment of the present invention relates to a method for producing melon seeds, the method comprising the following steps:

[0409] a) To induce growth in a melon plant containing at least one chromosome 5, wherein the at least one chromosome 5 has an introgression segment from the chromosome 5 of a ToLCNDV-ES resistant donor plant, the introgression segment containing a sequence from the donor plant between SNP_03 and SNP_04 (and / or containing one or more of SNP_01 to SNP_07, especially the donor SNPs of SNP_03 and / or SNP_04), and / or containing a QTL5rb, and / or containing a potential causal gene of QTL5rb (the causal gene may be a modified endogenous gene of the TIR-NBS-LRR protein-coding gene disclosed herein or a gene identified in a wild-type donor).

[0410] b) Harvest the fruits of the melon plants grown in step a).

[0411] c) Collect seeds from the fruit obtained in step b).

[0412] Seeds obtainable by the method for producing melon seeds according to the invention are also an embodiment of the invention. The seeds contain at least one copy of an introgression fragment comprising QTL5rb or at least one copy of QTL5rb or a potential causative gene for QTL5rb or an endogenous modification gene encoding the TIR-NBS-LRR protein disclosed herein.

[0413] Another embodiment of the present invention relates to a method for producing hybrid melon seeds, the method comprising the following steps

[0414] a) Provide a first inbred melon plant containing at least one chromosome 5, wherein the at least one chromosome 5 has an introgression fragment from chromosome 5 of a ToLCNDV-ES resistant donor plant, the introgression fragment containing a sequence from the donor plant between SNP_03 and SNP_04 (and / or containing one or more of SNP_01 to SNP_07, especially the donor SNPs of SNP_03 and / or SNP_04), and / or containing a QTL5rb, and / or containing a potential causal gene of QTL5rb (the causal gene may be a modified endogenous gene of the TIR-NBS-LRR protein-coding gene disclosed herein or a gene identified in a wild-type donor).

[0415] b) Provide a second-inbred melon plant with or without an introgression fragment from chromosome 5 of a ToLCNDV-ES resistant donor plant, the introgression fragment containing a sequence from the donor plant between SNP_03 and SNP_04 (and / or containing one or more of SNP_01 to SNP_07, especially the donor SNPs SNP_03 and / or SNP_04), and / or containing QTL5rb, and / or containing a potential causal gene of QTL5rb (the causal gene may be a modified endogenous gene of the TIR-NBS-LRR protein-coding gene disclosed herein or a gene identified in a wild-type donor).

[0416] c) Hybridize the plants provided in step a) with the plants provided in step b).

[0417] d) Select the seeds obtained from the hybridization in step c).

[0418] Therefore, in one aspect, the plant in step b) can be a second-inbred melon plant susceptible to ToLCNDV-ES, especially to strains that disrupt resistance, or it can be an inbred melon plant containing QTL5 (e.g., as in the variety Coliseo), whereby the hybrid melon contains both QTL5 and QTL5rb. This stacking of the two QTLs is particularly advantageous for conferring broad and durable resistance. Thus, in one aspect, cultivated melon plants are provided containing one QTL5rb (or the potential causal gene of QTL5rb, or its modified or mutated endogenous gene) and one QTL5 on each chromosome 5 of the genome. In one aspect, the cultivated melon plant is an F1 hybrid.

[0419] The term "inbred plant" or "inbred strain" in this invention shall mean a plant that has undergone several generations of self-pollination and is highly consistent in its genetic setting and phenotypic appearance.

[0420] As mentioned elsewhere in this document, when referring to a plant, plant part, cell or tissue, or fruit or seed containing a sequence between two SNPs, this in one respect covers the presence of a resistant donor nucleotide for one or more SNP nucleotides disclosed herein, particularly one or both of the mentioned SNPs and / or the SNP located between the mentioned SNPs, and optionally one or more of the remaining SNPs of the donor disclosed herein.

[0421] In a preferred embodiment of the invention, the inbred lines used in steps a) and b) of the method for producing hybrid melon seeds according to the invention have specific characteristics described as preferred and further embodiments of the plant according to the invention. The preferred and further embodiments described herein for the plant according to the invention are therefore applicable to the method for producing hybrid melon seeds according to the invention.

[0422] Hybrid seeds that can be obtained or obtained by the method for producing hybrid melon seeds according to the present invention are also an embodiment of the present invention.

[0423] Another embodiment of the present invention is a method for producing melon fruit, the method comprising the following steps

[0424] a) To induce growth in a melon plant containing at least one chromosome 5, wherein the at least one chromosome 5 has an introgression fragment from the chromosome 5 of a ToLCNDV-ES resistant donor plant, the introgression fragment containing a sequence between SNP_03 and SNP_04 of the donor plant and / or containing QTL5rb, and / or containing a potential causal gene of QTL5rb (the causal gene may be a modified endogenous gene of the TIR-NBS-LRR protein-coding gene disclosed herein or a gene identified in a wild-type donor).

[0425] b) Harvest the fruit produced by the plants grown in step a).

[0426] The term "fruit" in its botanical sense is generally understood as the seed-bearing structure that develops from the ovary of an angiosperm flower.

[0427] The melon fruit obtained by the method for producing melon fruit according to the present invention is also an embodiment of the present invention.

[0428] As described herein, melon donor plants resistant to ToLCNDV-ES (especially resistance-destroying strains) can be identified by means of SNP markers, particularly one or all of SNP_01, SNP_02, SNP_03, SNP_04, SNP_05, SNP_06 and SNP_07 disclosed herein and / or by the presence of causal genes of QTL5rb.

[0429] Therefore, in one aspect, the present invention enables those skilled in the art, for the first time, to identify donor plants from which introgression fragments conferring ToLCNDV-ES resistance to melon plants can be transferred to recurrent melon plants or to plants containing QTL5s that do not confer resistance to destructive strains. Thus, QTL5s can be replaced by QTL5rbs in melon plants, such as in the variety Coliseo F1 (Nunhems).

[0430] Melon donor plants can also be de novo produced by modifying endogenous genes, for example, using gene editing or mutagenesis methods, so that the endogenous genes encode the resistance TIR-NBS-LRR protein at a locus on chromosome 5.

[0431] Therefore, another embodiment of the invention relates to the use of one or more or all of SNP_01, SNP_02, SNP_03, SNP_04, SNP_05, SNP_06, and SNP_07 and / or potential causal genes of QTL5rb (e.g., any sequences provided herein, such as SEQ ID NO: 1 to 7, SEQ ID NO: 8, 11, or 12, or portions thereof) for the identification of ToLCNDV-ES resistant melon plants or parts thereof (such as cells, fruits, leaves), particularly melon plants or parts thereof resistant to strains that destroy resistance. Preferably, this use relates to the identification of ToLCNDV-ES resistant donor melon plants and / or recurrent melon plants, and also to the identification of breeding lines, cultivars, or varieties containing, for example, QTL5rb derived from seeds preserved herein or from another wild donor, or containing an endogenous modifying gene encoding a TIR-NBS-LRR resistance protein.

[0432] Another implementation is the use of one or more or all of SNP_01, SNP_02, SNP_03, SNP_04, SNP_05, SNP_06 or SNP_07 and / or QTL5rb and / or potential causal genes of QTL5rb (e.g., any sequence provided herein, such as SEQ ID NO: 1 to 7, SEQ ID NO: 8, 11 or 12, or portions thereof) to infiltrate ToLCNDV-ES resistance (especially resistance to strains that destroy resistance) into ToLCNDV-ES susceptible melon plants and / or into plants lacking QTL5rb (e.g., containing QTL5 at that locus), especially in cultivated melon lines or varieties.

[0433] Furthermore, one embodiment of the present invention is the use of one or more or all of SNP_01, SNP_02, SNP_03, SNP_04, SNP_05, SNP_06 or SNP_07 and / or QTL5rb and / or potential causal genes of QTL5rb (e.g., any sequence provided herein, such as SEQ ID NO: 1 to 7, SEQ ID NO: 8, 11 or 12, or portions thereof) in the cultivation of ToLCNDV-ES resistant melon plants (especially resistance to strains that destroy resistance).

[0434] A method is also provided for screening plants or plant materials or DNA derived therefrom for the presence of a fragment on chromosome 5 that confers TolCNDV-ES resistance. This method includes the following steps:

[0435] - Screen genomic DNA for one or more SNP haplotypes or genotypes and / or QTL5rb and / or potential causal genes in SNP_01, SNP_02, SNP_03, SNP_04, SNP_05, SNP_06 and SNP_07;

[0436] - and optionally, plants or plant materials containing one or more of the resistance donor genotypes of SNP_01, SNP_02, SNP_03, SNP_04, SNP_05, SNP_06 and SNP_07 and / or containing QTL5rb and / or containing the potential causal gene of QTL5rb.

[0437] In such screening and / or selection methods, any SNP markers and / or sequences or portions thereof provided herein may be used, such as SEQ ID NO: 1 to 7, SEQ ID NO: 8, 11 or 12 or portions thereof.

[0438] A method for producing cultivated melon plants containing an introgression fragment on chromosome 5 is also provided, wherein the introgression fragment comprises the ToLCNDV-ES QTL, the method comprising:

[0439] a) A first cultivated melon plant susceptible to ToLCNDV-ES (especially to destructive ToLCNDV-ES strains) is crossed with a second wild melon plant resistant to ToLCNDV-ES (especially to destructive ToLCNDV-ES strains), wherein the second melon plant contains the CC or CX genotype of SNP_01 at nucleotide 81 of SEQ ID NO: 1, and / or the CC or CX genotype of SNP_02 at nucleotide 242 of SEQ ID NO: 2 (or the GG or GX genotype when referring to the complementary strand), and / or the GG or GX genotype of SNP_03 at nucleotide 81 of SEQ ID NO: 3, and / or the AA or AX genotype of SNP_04 at nucleotide 101 of SEQ ID NO: 4 (or the TT or TX genotype when referring to the complementary strand), and / or the TT or TX genotype of SNP_05 at nucleotide 101 of SEQ ID NO: 5 (or the AA or AX genotype when referring to the complementary strand) and / or the AA or CX genotype of SNP_05 at nucleotide 101 of SEQ ID NO: 5. The AA or AX genotype of SNP_06 at nucleotide 101 of ID NO: 6 (or the TT or TX genotype when referring to the complementary strand), and / or the CC or CX genotype of SNP_07 in SEQ ID NO: 7; and / or wherein the second wild melon plant contains a gene encoding the protein of SEQ ID NO: 8 or a gene encoding a protein having at least 96% sequence identity with SEQ ID NO: 8, and wherein said protein preferably contains at least one of the following amino acids, preferably at least two, three or all four amino acids:

[0440] - Nine serines (S) at equivalent positions in amino acids 10 to 18 of SEQ ID NO: 8, or in a sequence having at least 96% sequence identity with SEQ ID NO: 8.

[0441] - Methionine (M) at amino acid number 484 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0442] - Lysine (K) at amino acid number 629 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0443] Histidine (H) or aspartic acid (D) at amino acid number 679 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0444] And / or one of the second wild melon plants contains

[0445] - The nucleotide sequence of SEQ ID NO: 11 or a sequence having at least 95%, 96%, 97%, 98% or 99% sequence identity with SEQ ID NO: 11;

[0446] b) Collect F1 seeds from the hybridization and backcross the F1 plants with a first melon plant to produce a backcross (BC1) population, or self-pollinate the F1 plants once or multiple times to produce an F2 or F3 population, and optionally self-pollinate the backcross population to produce a BC1S1 population.

[0447] The F2, F3, BC1, or BC1S1 plants described herein contain the CC or CX genotype of SNP_01 at nucleotide 81 of SEQ ID NO: 1, and / or the CC or CX genotype of SNP_02 at nucleotide 242 of SEQ ID NO: 2 (or the GG or GX genotype when referring to the complementary strand), and / or the GG or GX genotype of SNP_03 at nucleotide 81 of SEQ ID NO: 3, and / or the AA or AX genotype of SNP_04 at nucleotide 101 of SEQ ID NO: 4 (or the TT or TX genotype when referring to the complementary strand), and / or the TT or TX genotype of SNP_05 at nucleotide 101 of SEQ ID NO: 5 (or the AA or AX genotype when referring to the complementary strand), and / or the AA or AX genotype of SNP_06 at nucleotide 101 of SEQ ID NO: 6 (or the TT or TX genotype when referring to the complementary strand), and / or in SEQ ID NO: The CC or CX genotype of SNP_07 in 7;

[0448] And / or the F2, F3, BC1, or BC1S1 plant contains a gene encoding the protein of SEQ ID NO: 8 or a gene encoding a protein having at least 96% sequence identity with SEQ ID NO: 8, and the protein preferably contains at least one of the following amino acids, preferably at least two, three, or all four amino acids:

[0449] -Amino acids 10 to 18 of S0045Q ID NO: 8, or nine serines (S) at equivalent positions in a sequence having at least 96% sequence identity with SEQ ID NO: 8.

[0450] - Methionine (M) at amino acid number 484 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0451] Lysine (K) at amino acid number 629 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0452] - Histidine (H) or aspartic acid (D) at amino acid number 679 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0453] And / or said F2, F3, BC1 or BC1S1 plants contain

[0454] - The nucleotide sequence of SEQ ID NO: 11 or a sequence having at least 95%, 96%, 97%, 98% or 99% sequence identity with SEQ ID NO: 11.

[0455] A method is also provided for identifying or detecting cultivated melon plants containing an introgression fragment on chromosome 5, wherein the introgression fragment contains the ToLCNDV-ES resistance allele, the method comprising:

[0456] a) Screening of melon plants using the following assays: Molecular marker assays detecting at least one SNP marker selected from the group consisting of: SNP_01 of SEQ ID NO: 1, SNP_02 of SEQ ID NO: 2, SNP_03 of SEQ ID NO: 3, SNP_04 of SEQ ID NO: 4, SNP_05 of SEQ ID NO: 5, SNP_06 of SEQ ID NO: 6, and / or SNP_07 of SEQ ID NO: 7; and / or molecular assays (e.g., PCR, sequencing, molecular marker assays, etc.) detecting the presence of a gene encoding the protein of SEQ ID NO: 8 or a gene encoding a protein having at least 96% sequence identity with SEQ ID NO: 8, wherein said protein preferably contains at least one of the following amino acids, preferably at least two, three, or all four:

[0457] - Nine serines (S) at equivalent positions in amino acids 10 to 18 of SEQ ID NO: 8, or in a sequence having at least 96% sequence identity with SEQ ID NO: 8.

[0458] - Methionine (M) at amino acid number 484 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0459] - Lysine (K) at amino acid number 629 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0460] - Histidine (H) or aspartic acid (D) at amino acid number 679 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0461] And / or molecular assays (e.g., PCR, sequencing, molecular marker assays, etc.) to detect the presence of the following nucleotide sequences.

[0462] - The nucleotide sequence of SEQ ID NO: 11 or a sequence having at least 95%, 96%, 97%, 98%, or 99% sequence identity with SEQ ID NO: 11.

[0463] as well as

[0464] b) Identify and / or select plants containing the following: the CC or CX genotype of SNP_01 at nucleotide 81 of SEQ ID NO: 1, and / or the CC or CX genotype of SNP_02 at nucleotide 242 of SEQ ID NO: 2 (or the GG or GX genotype when referring to the complementary strand), and / or the GG or GX genotype of SNP_03 at nucleotide 81 of SEQ ID NO: 3, and / or the AA or AX genotype of SNP_04 at nucleotide 101 of SEQ ID NO: 4 (or the TT or TX genotype when referring to the complementary strand), and / or the TT or TX genotype of SNP_05 at nucleotide 101 of SEQ ID NO: 5 (or the AA or AX genotype when referring to the complementary strand) and / or the AA or AX genotype of SNP_06 at nucleotide 101 of SEQ ID NO: 6 (or the TT or TX genotype when referring to the complementary strand), and / or the CC or CX genotype of SNP_07 in SEQ ID NO: 7;

[0465] And / or identify and / or select plants containing a gene encoding the protein of SEQ ID NO: 8 or a gene encoding a protein having at least 96% sequence identity with SEQ ID NO: 8, wherein said protein preferably contains at least one of the following amino acids, preferably at least two, three, or all four amino acids:

[0466] - Nine serines (S) at equivalent positions in amino acids 10 to 18 of SEQ ID NO: 8, or in a sequence having at least 96% sequence identity with SEQ ID NO: 8.

[0467] - Methionine (M) at amino acid number 484 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0468] - Lysine (K) at amino acid number 629 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0469] - Histidine (H) or aspartic acid (D) at amino acid number 679 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0470] And / or identify and / or select plants containing the following:

[0471] - The nucleotide sequence of SEQ ID NO: 11 or a sequence having at least 95%, 96%, 97%, 98% or 99% sequence identity with SEQ ID NO: 11.

[0472] A method is provided for generating F1 melon hybrids containing the ToLCNDV-ES resistance phenotype (especially resistance to strains that destroy resistance), the method comprising:

[0473] a) Crossing a first inbred melon plant containing at least one recombinant chromosome 5 with a second inbred melon plant having or lacking said at least one recombinant chromosome 5 or lacking QTL5rb (but optionally replaced with QTL5), wherein the recombinant chromosome 5 contains an introgression fragment conferring ToLCNDV-ES resistance to the first inbred melon plant when present in homozygous or heterozygous form (i.e., containing a homozygous or heterozygous QTL5rb), and wherein said introgression fragment originates from a wild plant of the melon species, and

[0474] b) Collect F1 hybrid seeds from the hybridization.

[0475] Further, a method for producing melon plants resistant to ToLCNDV-ES on chromosome 5 is included, the method comprising:

[0476] a) Screening of one or more wild melon germplasms using the following assays: Detection of at least one SNP marker selected from the group consisting of: molecular marker assays of SNP_01 in SEQ ID NO: 1, SNP_02 in SEQ ID NO: 2, SNP_03 in SEQ ID NO: 3, SNP_04 in SEQ ID NO: 4, SNP_05 in SEQ ID NO: 5, SNP_06 in SEQ ID NO: 6, and / or SNP_07 in SEQ ID NO: 7; and / or molecular assays (e.g., PCR, sequencing, molecular marker assays, etc.) detecting the presence of a gene encoding the protein of SEQ ID NO: 8 or a gene encoding a protein having at least 96% sequence identity with SEQ ID NO: 8, wherein said protein preferably contains at least one of the following amino acids, preferably at least two, three, or all four amino acids.

[0477] - Nine serines (S) at equivalent positions in amino acids 10 to 18 of SEQ ID NO: 8, or in a sequence having at least 96% sequence identity with SEQ ID NO: 8.

[0478] - Methionine (M) at amino acid number 484 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0479] - Lysine (K) at amino acid number 629 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0480] - Histidine (H) or aspartic acid (D) at amino acid number 679 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0481] And / or molecular assays (e.g., PCR, sequencing, molecular marker assays, etc.) to detect the presence of the following nucleotide sequences.

[0482] - The nucleotide sequence of SEQ ID NO: 11 or a sequence having at least 95%, 96%, 97%, 98% or 99% sequence identity with SEQ ID NO: 11;

[0483] b) Identification and / or selection

[0484] The wild melon plant includes the following: the CC or CX genotype of SNP_01 at nucleotide 81 of SEQ ID NO: 1, and / or the CC or CX genotype of SNP_02 at nucleotide 242 of SEQ ID NO: 2 (or the GG or GX genotype when referring to the complementary strand), and / or the GG or GX genotype of SNP_03 at nucleotide 81 of SEQ ID NO: 3, and / or the AA or AX genotype of SNP_04 at nucleotide 101 of SEQ ID NO: 4 (or the TT or TX genotype when referring to the complementary strand), and / or the TT or TX genotype of SNP_05 at nucleotide 101 of SEQ ID NO: 5 (or the AA or AX genotype when referring to the complementary strand), and / or the AA or AX genotype of SNP_06 at nucleotide 101 of SEQ ID NO: 6 (or the TT or TX genotype when referring to the complementary strand), and / or the CC or CX genotype of SNP_07 in SEQ ID NO: 7;

[0485] And / or identify and / or select wild melon plants containing a gene encoding the protein of SEQ ID NO: 8 or a gene encoding a protein having at least 96% sequence identity with SEQ ID NO: 8, wherein said protein preferably contains at least one of the following amino acids, preferably at least two, three, or all four amino acids:

[0486] - Nine serines (S) at equivalent positions in amino acids 10 to 18 of SEQ ID NO: 8, or in a sequence having at least 96% sequence identity with SEQ ID NO: 8.

[0487] - Methionine (M) at amino acid number 484 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0488] - Lysine (K) at amino acid number 629 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0489] - Histidine (H) or aspartic acid (D) at amino acid number 679 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0490] And / or identify and / or select wild melon plants containing the following:

[0491] The nucleotide sequence of SEQ ID NO: 11 or a sequence having at least 95%, 96%, 97%, 98%, or 99% sequence identity with SEQ ID NO: 11.

[0492] c) Optionally, in resistance assays, especially using strains that destroy resistance, to confirm ToLCNDV-ES resistance;

[0493] d) and optionally, infiltrating the ToLCNDV-ES resistance from the wild germplasm into cultivated melon plants.

[0494] A cultivated melon plant or a portion thereof containing recombinant chromosome 5, wherein the recombinant chromosome 5 contains an introgression fragment that confers ToLCNDV-ES resistance to the melon plant when present in homozygous or heterozygous form, and wherein said introgression fragment contains one or more single nucleotide polymorphism (SNP) markers from the following group:

[0495] The CC or CX genotype of SNP_01 at nucleotide 81 of SEQ ID NO: 1, and / or the CC or CX genotype of SNP_02 at nucleotide 242 of SEQ ID NO: 2 (or the GG or GX genotype when referring to the complementary strand), and / or the GG or GX genotype of SNP_03 at nucleotide 81 of SEQ ID NO: 3, and / or the AA or AX genotype of SNP_04 at nucleotide 101 of SEQ ID NO: 4 (or the TT or TX genotype when referring to the complementary strand), and / or the TT or TX genotype of SNP_05 at nucleotide 101 of SEQ ID NO: 5 (or the AA or AX genotype when referring to the complementary strand), and / or the AA or AX genotype of SNP_06 at nucleotide 101 of SEQ ID NO: 6 (or the TT or TX genotype when referring to the complementary strand), and / or the CC or CX genotype of SNP_07 in SEQ ID NO: 7.

[0496] And optionally, in one aspect, the infiltrated fragment comes from wild plants of a melon species, and the average ToLCNDV-ES disease score of the wild plants is at least 7.7, 7.8, 7.9, 8.0, preferably at least 9.0, according to a scale from 1 = plant death to 9 = asymptomatic.

[0497] In one aspect, the cultivated melon plant contains at least the GG or GX genotype of SNP_03 at nucleotide 81 of SEQ ID NO: 3 and / or the AA or AX genotype of SNP_04 at nucleotide 101 of SEQ ID NO: 4 (or the TT or TX genotype when referring to the complementary strand).

[0498] In one respect, the ToLCNDV-ES resistance QTL5rb or an infiltrated fragment containing QTL5rb is available from / may be available from / is present in seeds or their progeny containing a representative sample deposited under accession number NCIMB44139 (thereby the progeny retain ToLCNDV-ES resistance).

[0499] A cultivated melon plant or a portion thereof comprising recombinant chromosome 5, the recombinant chromosome 5 containing an introgression fragment that confers ToLCNDV-ES resistance to the melon plant when present in homozygous or heterozygous form, and said introgression fragment comprising:

[0500] The gene encoding the protein of SEQ ID NO: 8 or the gene encoding a protein having at least 96% sequence identity with SEQ ID NO: 8, wherein said protein preferably contains at least one of the following amino acids, preferably at least two, three or all four amino acids:

[0501] - Nine serines (S) at equivalent positions in amino acids 10 to 18 of SEQ ID NO: 8, or in a sequence having at least 96% sequence identity with SEQ ID NO: 8.

[0502] - Methionine (M) at amino acid number 484 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0503] - Lysine (K) at amino acid number 629 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0504] - Histidine (H) or aspartic acid (D) at amino acid number 679 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8;

[0505] And / or the nucleotide sequence of SEQ ID NO: 11 or having at least 95%, 96%, 97%, 98% or 99% sequence identity with SEQ ID NO: 11, preferably a sequence encoding the above-mentioned protein,

[0506] And optionally, in one aspect, the infiltrated fragment comes from wild plants of the melon species, and the average ToLCNDV-ES disease score of the wild plants is at least 7.7, 7.8, 7.9, 8.0, 8.5, preferably at least 9.0, according to a scale from 1 = plant death to 9 = asymptomatic.

[0507] In one respect, the ToLCNDV-ES resistance QTL5rb or an infiltrated fragment containing QTL5rb is available from / may be available from / is present in seeds or their progeny containing a representative sample deposited under accession number NCIMB44139 (thereby the progeny retain ToLCNDV-ES resistance).

[0508] In another aspect, melon plants (or plant parts, such as cells) containing QTL5rb can be de novo produced by introducing one or more modifications into the same locus in the susceptibility gene on chromosome 5 through random or targeted mutagenesis to convert the susceptible TIR-NBS-LRR protein into the resistant TIR-NBS-LRR protein.

[0509] In susceptible cultivated melons, a locus on chromosome 5 contains a gene encoding the protein of SEQ ID NO: 10 (or a susceptible protein having at least 98%, 98.5%, 99%, 99.5%, or 99.8% sequence identity with SEQ ID NO: 10). This TIR-NBS-LRR protein is referred to herein as the "susceptible protein." In one aspect, it can be altered into a "resistant protein" by changing one or more (or all) of the four amino acid differences required to make the protein a "resistant protein" (i.e., conferring resistance to ToLCNDV-ES). See also Figure 2 The four differences between the susceptible and resistant proteins are numbered 1 to 4.

[0510] Random mutagenesis, followed by selection (e.g., using tilling) of plants containing mutant alleles, and / or targeted mutagenesis can be used to introduce one or more of variations 1 to 4 into plants containing genes encoding, for example, a susceptibility protein of SEQ ID NO: 10 or a susceptibility protein having at least 98%, 98.5%, 99%, 99.5%, or 99.8% sequence identity with SEQ ID NO: 10.

[0511] Therefore, in one aspect, a melon plant or a portion thereof is provided, the melon plant or the portion thereof comprising a modified allele of a gene encoding the TIR-NBS-LRR protein of SEQ ID NO: 10 (or a susceptibility protein having at least 98%, 98.5%, 99.8%, 99%, 99.2%, 99.5%, or 99.8% identity with SEQ ID NO: 10), wherein the modified allele, when present in a homozygous or heterozygous form, confers ToLCNDV-ES resistance to the melon plant, preferably ToLCNDV-ES resistance against a resistant strain, and wherein the modified allele encodes a protein having at least 96% sequence identity with SEQ ID NO: 8, and wherein the protein preferably comprises at least one of the following amino acids, preferably at least two, three, or all four amino acids:

[0512] - Nine serine residues (S) at equivalent positions in amino acids 10 to 18 of SEQ ID NO: 8, or in a sequence having at least 96% sequence identity with SEQ ID NO: 8; (change 1 compared to the susceptibility protein)

[0513] - Methionine (M) at amino acid number 484 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8; (change 2 compared to the susceptibility protein)

[0514] - Lysine (K) at amino acid number 629 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8; (change 3 compared to the susceptible protein)

[0515] - Histidine (H) or aspartic acid (D) at amino acid number 679 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8; (change 4 compared to the susceptibility protein).

[0516] Therefore, the modified allele encoding the susceptibility protein of SEQ ID NO: 10, or the susceptibility protein of SEQ ID NO: 10 with at least 98%, 98.5%, 99%, 99.5%, or 99.8% sequence identity, comprises one or all of the following four amino acid changes:

[0517] 1. At the beginning of the protein, two S (serine) residues are inserted after amino acid E9 (glutamic acid number 9); that is, the protein contains 9 serine residues (S) after E9, and amino acids 10 to 18 are serine residues in proteins having at least 96%, 97%, 98%, or 99% sequence identity with the protein of SEQ ID NO: 8 (while SEQ ID NO: 10 contains only 7 serine residues, i.e., amino acids 10 to 16); see also Figure 2 Change number 1;

[0518] 2. At the equivalent position in a protein having at least 96%, 97%, 98%, or 99% sequence identity with the protein of SEQ ID NO: 8, amino acid V483 (valine 483 of SEQ ID NO: 10) is changed to M (methionine); in SEQ ID NO: 8, the equivalent position is M484; see also Figure 2 Change number 2;

[0519] 3. In proteins having at least 96%, 97%, 98%, or 99% sequence identity with the protein of SEQ ID NO: 8, amino acid E628 (glutamic acid 628 of SEQ ID NO: 10) is changed to K (lysine); in SEQ ID NO: 8, the equivalent position is K629; see also Figure 2 Change number 3;

[0520] 4. At the equivalent position in a protein having at least 96%, 97%, 98%, or 99% sequence identity with the protein of SEQ ID NO: 8, amino acid Y678 (tyrosine 678 of SEQ ID NO: 10) is changed to H (histidine); in SEQ ID NO: 8, the equivalent position is H679, see [link to relevant documentation]. Figure 2 Change 4; or at the equivalent position in a protein having at least 96%, 97%, 98%, or 99% sequence identity with the protein of SEQ ID NO: 8, amino acid Y678 (tyrosine 678 of SEQ ID NO: 10) is changed to D (aspartic acid); in SEQ ID NO: 8, the equivalent position is D679, see [link to SEQ ID NO: 8]. Figure 1 or Figure 2 Change number 4.

[0521] In one aspect, the endogenous genome sequence encoding the susceptibility protein is modified in cultivated melon plants so that the genome sequence encodes the resistance protein instead of the susceptibility protein, thus encoding a protein containing one or more of the four variations listed above.

[0522] For example, Figure 4This demonstrates how to modify SEQ ID NO: 36, which encodes the susceptibility protein of SEQ ID NO: 10, to convert it into a resistance protein.

[0523] For example, change 1 involves inserting the codon TCT, which encodes serine, twice, resulting in two serine residues in the resistant protein. Change 2 changes the codon GTG (valine, V) to ATG (methionine, M), change 3 changes the codon GAG (glutamate, E) to AAG (lysine, K), and change 4 changes the codon TAT (tyrosine, Y) to CAT (histidine, H).

[0524] Therefore, in one aspect, a cultivated melon plant or a portion thereof (e.g., cell or tissue or fruit or seed) is provided, the cultivated melon plant or the portion thereof containing an allele encoding a TIR-NBS-LRR protein, which, when present in a homozygous or heterozygous form, confers ToLCNDV-ES resistance to the melon plant, preferably ToLCNDV-ES resistance against a resistant strain, and wherein said allele encodes a protein having at least or 96%, 97%, 98% or 99% sequence identity with SEQ ID NO: 8, and wherein said protein preferably contains at least one of the following amino acids, preferably at least two, three or all four amino acids:

[0525] - Nine serine residues (S) at equivalent positions in amino acids 10 to 18 of SEQ ID NO: 8, or in a sequence having at least 96% sequence identity with SEQ ID NO: 8; (change 1 compared to the susceptibility protein)

[0526] - Methionine (M) at amino acid number 484 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8; (change 2 compared to the susceptibility protein)

[0527] - Lysine (K) at amino acid number 629 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8; (change 3 compared to the susceptible protein)

[0528] - Histidine (H) or aspartic acid (D) at amino acid number 679 of SEQ ID NO: 8 or at an equivalent position in a sequence having at least 96% sequence identity with SEQ ID NO: 8; (change 4 compared to the susceptibility protein).

[0529] Therefore, in one aspect, cultivated melon plants, seeds, or plant parts may contain mutant or modified TIR-NBS-LRR alleles, wherein the mutant or modified alleles (also referred to herein as modified alleles) are generated by random mutagenesis or targeted mutagenesis (such as CRISPR-based methods). Random mutagenesis can be, for example, chemically induced (e.g., EMS treatment) or radiation-induced mutagenesis or other methods by which mutations are randomly induced in the genome, and plants or plant parts containing mutations of the endogenous TIR-NBS-LRR gene can then be screened and identified. Targeted mutagenesis is a method of specifically introducing mutations into a target gene such as the TIR-NBS-LRR gene using, for example, Crispr-Cas9 or Crispr-CpfI or other known methods. It should be noted that, using such methods, the mutant alleles described herein can be generated without excessive burden.

[0530] When this article refers to melon plants, in one respect, it covers a seed from which the plant can grow, namely, an embryo in the seed that can contain at least one (or two) copy of the described mutant allele.

[0531] In one aspect, the melon plant containing the mutant allele or modified allele is not produced solely through a fundamentally biological process; this means that the mutant allele was produced somewhere through human intervention. The patent covers plants containing mutant alleles even if the plant itself was produced solely through hybridization and selection, if such human-generated mutant alleles were transferred from one plant to another through hybridization and selection. Preferably, in one aspect, the plant is not transgenic, and any constructs used to modify the endogenous gene, for example, in the case of targeted gene editing, have been removed from the genome. Furthermore, in another aspect, the plant is not transgenic because the mutant allele has not been introduced from outside and has not been integrated anywhere in the plant genome using plant transformation technology, but the mutant allele is an endogenous TIR-NBS-LRR allele that has been mutated or modified (using, for example, targeted or random mutagenesis) at the locus on chromosome 5 in the genome where the TIR-NBS-LRR allele is located.

[0532] In one respect, particularly with regard to the European Patent Convention, the plant according to the invention "is not obtained solely by essentially biological means".

[0533] Therefore, in one respect, the endogenous mutant or modified TIR-NBS-LRR allele is an induced mutant allele.

[0534] In one respect, plants do not contain any chimeric constructs in their genomes.

[0535] Detection and / or selection methods

[0536] A method is provided for selecting melon plants (especially cultivated melons) or plant parts containing chromosome 5, wherein the chromosome 5 containing the introgression fragment comprises a quantitative trait locus (QTL) named QTL5rb between a cytosine of SNP_01 at nucleotide 81 of SEQ ID NO: 1 (nucleotide 25.456.098) corresponding to chromosome 5 of the melon genome and a cytosine of SNP_07 at nucleotide 81 of SEQ ID NO: 7 (nucleotide 26.390.451) corresponding to chromosome 5 of the melon genome, the QTL5rb conferring resistance to New Delhi Tomato Leaf Curl Virus (ToLCNDV-ES) to the melon plant when present in homozygous or heterozygous form, the method comprising selecting plants or plant parts containing one or all of the following single nucleotide polymorphism (SNP) markers:

[0537] a) Cytosine at nucleotide 242 of SEQ ID NO: 2, or at an equivalent nucleotide in a sequence having at least 95% sequence identity with SEQ ID NO: 2 (or guanine at nucleotide 242 of the complementary sequence of SEQ ID NO: 2, or at an equivalent nucleotide in a sequence having at least 95% sequence identity with the complementary sequence of SEQ ID NO: 2);

[0538] b) Guanine at nucleotide 81 of SEQ ID NO: 3, or at an equivalent nucleotide in a sequence having at least 95% sequence identity with SEQ ID NO: 3;

[0539] c) Adenine at nucleotide 101 of SEQ ID NO: 4, or at an equivalent nucleotide in a sequence having at least 95% sequence identity with SEQ ID NO: 4 (or thymine at nucleotide 101 of the complementary sequence of SEQ ID NO: 4, or at an equivalent nucleotide in a sequence having at least 95% sequence identity with the complementary sequence of SEQ ID NO: 4).

[0540] d) Thymine at nucleotide 101 of SEQ ID NO: 5, or at an equivalent nucleotide in a sequence having at least 95% sequence identity with SEQ ID NO: 5 (or adenine at nucleotide 101 of the complementary sequence of SEQ ID NO: 5, or at an equivalent nucleotide in a sequence having at least 95% sequence identity with the complementary sequence of SEQ ID NO: 5).

[0541] e) Adenine at nucleotide 101 of SEQ ID NO: 6 or at an equivalent nucleotide in a sequence having at least 95% sequence identity with SEQ ID NO: 6 (or thymine at nucleotide 101 of the complementary sequence of SEQ ID NO: 6 or at an equivalent nucleotide in a sequence having at least 95% sequence identity with the complementary sequence of SEQ ID NO: 6).

[0542] In another aspect, a method is provided for selecting melon plants or plant parts, wherein the plant contains chromosome 5, and the chromosome 5 containing the introgression fragment contains a quantitative trait locus (QTL) named QTL5rb between a cytosine of SNP_01 at nucleotide 81 of SEQ ID NO: 1 (nucleotide 25.456.098) corresponding to chromosome 5 of the melon genome and a cytosine of SNP_07 at nucleotide 81 of SEQ ID NO: 7 (nucleotide 26.390.451) corresponding to chromosome 5 of the melon genome, the QTL5rb conferring resistance to New Delhi Tomato Leaf Curl Virus (ToLCNDV-ES) to the melon plant when present in homozygous or heterozygous form, the method comprising selecting plants or plant parts containing one or all of the following single nucleotide polymorphism (SNP) markers:

[0543] a) Cytosine at nucleotide 81 of SEQ ID NO: 1, or at an equivalent nucleotide in a sequence having at least 95% sequence identity with SEQ ID NO: 1;

[0544] b) Cytosine at nucleotide 242 of SEQ ID NO: 2, or at an equivalent nucleotide in a sequence having at least 95% sequence identity with SEQ ID NO: 2; (or guanine at nucleotide 242 of the complementary sequence of SEQ ID NO: 2, or at an equivalent nucleotide in a sequence having at least 95% sequence identity with the complementary sequence of SEQ ID NO: 2).

[0545] c) Guanine at nucleotide 81 of SEQ ID NO: 3, or at an equivalent nucleotide in a sequence having at least 95% sequence identity with SEQ ID NO: 3;

[0546] d) Adenine at nucleotide 101 of SEQ ID NO: 4, or at an equivalent nucleotide in a sequence having at least 95% sequence identity with SEQ ID NO: 4; (or thymine at nucleotide 101 of the complementary sequence of SEQ ID NO: 4, or at an equivalent nucleotide in a sequence having at least 95% sequence identity with the complementary sequence of SEQ ID NO: 4).

[0547] e) Thymine at nucleotide 101 of SEQ ID NO: 5, or at an equivalent nucleotide in a sequence having at least 95% sequence identity with SEQ ID NO: 5; (or adenine at nucleotide 101 of the complementary sequence of SEQ ID NO: 5, or at an equivalent nucleotide in a sequence having at least 95% sequence identity with the complementary sequence of SEQ ID NO: 5).

[0548] f) Adenine at nucleotide 101 of SEQ ID NO: 6 or at an equivalent nucleotide in a sequence having at least 95% sequence identity with SEQ ID NO: 6 (or thymine at nucleotide 101 of the complementary sequence of SEQ ID NO: 6 or at an equivalent nucleotide in a sequence having at least 95% sequence identity with the complementary sequence of SEQ ID NO: 6).

[0549] g) Cytosine at nucleotide 101 of SEQ ID NO: 7 or at an equivalent nucleotide in a sequence having at least 95% sequence identity with SEQ ID NO: 7.

[0550] For the choice, for example, the KASP assay or other genotyping assays can be used, see examples.

[0551] Therefore, in one aspect, a genotyping assay is provided for genotyping melon plants, seeds, plant parts, cells, or tissues, the assay comprising the following steps:

[0552] a) Provide genomic DNA of one or more melon plants or plant populations (e.g., breeding populations, F2 populations, backcross populations, etc.), and

[0553] b) Genotyping assays that detect the presence of one or more of the following biomarkers:

[0554] - Cytosine at nucleotide 242 of SEQ ID NO: 2, or at an equivalent nucleotide in a sequence having at least 95% sequence identity with SEQ ID NO: 2; (or guanine at nucleotide 242 of the complementary sequence of SEQ ID NO: 2, or at an equivalent nucleotide in a sequence having at least 95% sequence identity with the complementary sequence of SEQ ID NO: 2).

[0555] - Guanine at nucleotide 81 of SEQ ID NO: 3, or at an equivalent nucleotide in a sequence having at least 95% sequence identity with SEQ ID NO: 3;

[0556] - Adenine at nucleotide 101 of SEQ ID NO: 4, or at an equivalent nucleotide in a sequence having at least 95% sequence identity with SEQ ID NO: 4; (or thymine at nucleotide 101 of the complementary sequence of SEQ ID NO: 4, or at an equivalent nucleotide in a sequence having at least 95% sequence identity with the complementary sequence of SEQ ID NO: 4).

[0557] - Thymine at nucleotide 101 of SEQ ID NO: 5, or at an equivalent nucleotide in a sequence having at least 95% sequence identity with SEQ ID NO: 5; (or adenine at nucleotide 101 of the complementary sequence of SEQ ID NO: 5, or at an equivalent nucleotide in a sequence having at least 95% sequence identity with the complementary sequence of SEQ ID NO: 5).

[0558] - Adenine at nucleotide 101 of SEQ ID NO: 6 or at an equivalent nucleotide in a sequence having at least 95% sequence identity with SEQ ID NO: 6 (or thymine at nucleotide 101 of the complementary sequence of SEQ ID NO: 6 or at an equivalent nucleotide in a sequence having at least 95% sequence identity with the complementary sequence of SEQ ID NO: 6).

[0559] as well as

[0560] c) Select a plant, seed, plant part, cell, or tissue that contains one or more SNP nucleotides from step b).

[0561] Melon plants can be wild melon germplasm or cultivated melon plants.

[0562] In another aspect, a genotyping assay is provided for genotyping melon plants, plant parts, cells, or tissues, the assay comprising the following steps:

[0563] a) Provide genomic DNA of one or more melon plants or plant populations (e.g., breeding populations, F2 populations, backcross populations, etc.), and

[0564] b) Genotyping tests to detect the presence of one or more of the following:

[0565] - Cytosine at nucleotide 242 of SEQ ID NO: 2, or at an equivalent nucleotide in a sequence having at least 95% sequence identity with SEQ ID NO: 2; (or guanine at nucleotide 242 of the complementary sequence of SEQ ID NO: 2, or at an equivalent nucleotide in a sequence having at least 95% sequence identity with the complementary sequence of SEQ ID NO: 2).

[0566] - Guanine at nucleotide 81 of SEQ ID NO: 3, or at an equivalent nucleotide in a sequence having at least 95% sequence identity with SEQ ID NO: 3;

[0567] - Adenine at nucleotide 101 of SEQ ID NO: 4, or at an equivalent nucleotide in a sequence having at least 95% sequence identity with SEQ ID NO: 4; (or thymine at nucleotide 101 of the complementary sequence of SEQ ID NO: 4, or at an equivalent nucleotide in a sequence having at least 95% sequence identity with the complementary sequence of SEQ ID NO: 4).

[0568] - Thymine at nucleotide 101 of SEQ ID NO: 5, or at an equivalent nucleotide in a sequence having at least 95% sequence identity with SEQ ID NO: 5; (or adenine at nucleotide 101 of the complementary sequence of SEQ ID NO: 5, or at an equivalent nucleotide in a sequence having at least 95% sequence identity with the complementary sequence of SEQ ID NO: 5).

[0569] - Adenine at nucleotide 101 of SEQ ID NO: 6 or at an equivalent nucleotide in a sequence having at least 95% sequence identity with SEQ ID NO: 6 (or thymine at nucleotide 101 of the complementary sequence of SEQ ID NO: 6 or at an equivalent nucleotide in a sequence having at least 95% sequence identity with the complementary sequence of SEQ ID NO: 6).

[0570] and optional

[0571] c) Select a plant, seed, plant part, cell, or tissue containing one or more SNP nucleotides in the homozygous form of step b).

[0572] Step a) may include isolating genomic DNA from the plant, seed, plant part, cell, or tissue to be analyzed in the genotyping assay. As is known in the art, crude DNA extraction methods can typically be used.

[0573] Step b) preferably includes biallelic genotyping, which utilizes allele-specific primers and / or allele-specific probes.

[0574] In another aspect, a method is provided for selecting melon plants containing chromosome 5, wherein the chromosome 5 containing an introgression fragment comprises a quantitative trait locus (QTL) named QTL5rb between a cytosine of SNP_01 at nucleotide 81 of SEQ ID NO: 1 (nucleotide 25.456.098) corresponding to chromosome 5 of the melon genome and a cytosine of SNP_07 at nucleotide 81 of SEQ ID NO: 7 (nucleotide 26.390.451) corresponding to chromosome 5 of the melon genome, the QTL5rb conferring resistance to New Delhi Tomato Leaf Curl Virus (ToLCNDV-ES) to the melon plant, the method comprising selecting a plant or plant portion containing a gene encoding a protein of SEQ ID NO: 8 or a gene encoding a protein having at least 96% sequence identity with SEQ ID NO: 8, the gene being located at SEQ ID NO: 25.956.264 (nucleotide 25.956.264) corresponding to chromosome 5 of the melon genome. The SNP_03 at nucleotide 81 of nucleotide 3 is between the SNP_04 at nucleotide 101 of nucleotide 26.040.072 of chromosome 5 of the melon genome, corresponding to SEQ ID NO: 4.

[0575] In one aspect of the above method, the gene encodes a protein having at least 96%, 97%, 98%, or 99% sequence identity with SEQ ID NO: 8, and contains one or more of the following amino acids:

[0576] a) Amino acids numbered 10 to 18 are serine (S).

[0577] b) The amino acid at the equivalent position of amino acid 484 in SEQ ID NO: 8 is methionine (M).

[0578] c) The amino acid at the equivalent position of amino acid 629 in SEQ ID NO: 8 is lysine (K).

[0579] d) The amino acid at the equivalent position of amino acid 679 in SEQ ID NO: 8 is either histidine (H) or aspartic acid (D).

[0580] Various methods can be used to select plants or plant parts that contain the mentioned genes. Selection can be based on genotyping methods (such as KASP assays), gene sequencing, mRNA (or cDNA) analysis, etc.

[0581] In another aspect, a method is provided for modifying an endogenous gene encoding the protein of SEQ ID NO: 10 (or a susceptibility protein having at least 98%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.7%, 98.8%, 98.9%, 99%, 99.5%, or 99.8% sequence identity with SEQ ID NO: 10) on chromosome 5 of melon by targeted gene editing or by mutagenesis, such as by radiation or chemical mutagenesis, thereby modifying said gene such that the protein encoded by SEQ ID NO: 8 having at least 96% sequence identity preferably comprises one or more of the following amino acid variations:

[0582] a) Amino acids numbered 10 to 18 are serine (S).

[0583] b) The amino acid at the equivalent position of amino acid 484 in SEQ ID NO: 8 is methionine (M).

[0584] c) The amino acid at the equivalent position of amino acid 629 in SEQ ID NO: 8 is lysine (K).

[0585] d) The amino acid at the equivalent position of amino acid 679 in SEQ ID NO: 8 is either histidine (H) or aspartic acid (D).

[0586] In this method, susceptible melon plants, seeds, or plant parts containing the TIR-NBS-LRR gene encoding the ToLCNDV-ES susceptibility protein, such as the protein in SEQ ID NO: 10, or the susceptibility protein having at least 98%, 98.1%, 98.2%, 98.3%, 98.4%, 98.5%, 98.6%, 98.7%, 98.8%, 98.9%, 99%, 99.5%, or 99.8% sequence identity with the susceptibility protein in SEQ ID NO: 10, are treated, for example, with a mutagen (e.g., radiation or chemical mutagen). Alternatively, the plant, seed, or plant part is provided for mutagenesis, and mutant alleles in the gene are selected.

[0587] Alternatively, targeted gene editing can be used to induce modifications in genes encoding susceptibility proteins. Thus, CRISPR-based methods, for example, can be used to modify the endogenous TIR-NBS-LRR gene on chromosome 5.

[0588] A method is provided for producing cultivated melon plants containing ToLCNDV-ES resistance, the method comprising the following steps:

[0589] a) Introducing random or targeted mutations (e.g., using a CRISPR-based approach) into one or more melon plants, plant parts, or seeds; or providing mutant plant or seed populations (e.g., TILLING populations, such as M2, M3, M4, or higher generations).

[0590] b) Select plants containing a mutant allele of the TIR-NBS-LRR gene on chromosome 5, wherein the mutant allele is modified such that the protein encoded by the allele having at least 96% sequence identity with SEQ ID NO: 8 preferably contains one or more of the following amino acid variations:

[0591] - Amino acids numbered 10 to 18 are serine (S).

[0592] - The amino acid at the equivalent position of amino acid 484 in SEQ ID NO: 8 is methionine (M).

[0593] The amino acid at the equivalent position of amino acid 629 in SEQ ID NO: 8 is lysine (K).

[0594] The amino acid at the equivalent position of amino acid 679 in SEQ ID NO: 8 is either histidine (H) or aspartic acid (D).

[0595] c) Optionally remove any transgenic constructs (e.g., CRISPR constructs) from the plant, and / or

[0596] d) Optionally generate plants with homozygous mutant alleles and analyze whether the plants are resistant to ToLCNDV-ES, especially the resistance-destroying ToLCNDV-ES strain.

[0597] This article also covers modified or mutated plants and plant parts obtained or available through any of the methods described above, from which seeds of such plants can be grown.

[0598] Genetically modified plants

[0599] In another aspect, a transgenic plant comprising a transgene encoding a resistance TIR-NBS-LRR protein, such as the protein of SEQ ID NO: 8 or a protein having at least 96%, 97%, 98%, or 99% sequence identity with SEQ ID NO: 8, wherein the protein preferably comprises one or more (or all) of the following amino acids:

[0600] a) Amino acids numbered 10 to 18 are serine (S).

[0601] b) The amino acid at the equivalent position of amino acid 484 in SEQ ID NO: 8 is methionine (M).

[0602] c) The amino acid at the equivalent position of amino acid 629 in SEQ ID NO: 8 is lysine (K).

[0603] d) The amino acid at the equivalent position of amino acid 679 in SEQ ID NO: 8 is either histidine (H) or aspartic acid (D).

[0604] Genetically modified plants can be any species; in one respect, it is melon, with a preference for cultivated melons.

[0605] The transgene can be inserted anywhere in the melon genome. Since the gene is dominant, it is not important that chromosome 5 contains the gene encoding the susceptibility TIR-NBS-LRR protein.

[0606] Seed Preservation Information

[0607] A representative sample of melon seeds from the ToLCNDV-ES resistance donor (named Melon ToLQTL5RB) was deposited by Nunhems B.V. on 20 April 2023 at NCIMB Ltd. (Ferguson Building, Craibstone Estate, Bucksburn Aberdeen, Scotland AB21 9YA, UK) under the Budapest Treaty, in accordance with the Expert Solution (EPC 2000, Rule 32(1)). The seeds are given accession number NCIMB 44139.

[0608] The applicant requests that, in accordance with the relevant regulations of EPC Rule 32(1) EPC or of a country or treaty with similar rules and regulations, samples of the biological material and any material derived therefrom be published only to a designated expert until the patent is granted, or until 20 years from the date of filing if the application is refused, withdrawn or deemed withdrawn.

[0609] During the pending period of this application, the depositary may be obtained by the person entitled to it upon request, as determined by the Director of the U.S. Patent and Trademark Office. Pursuant to 37 CFR § 1.808(b), all restrictions imposed by the depositary on the public availability of the depositary material will be irrevocably lifted upon patent grant. The depositary will remain for 30 years, or until 5 years after the most recent request, or for the duration of the patent, whichever is longer, and will be replaced if it becomes inactive during that period. The applicant does not waive any rights granted under this patent application or the Plant Variety Protection Act (7 USC 2321 ef)

[0610] Sequence description

[0611] In the SEQ ID NO shown in the sequence listing, the characters other than G (guanine), A (adenine), T (thymine), and C (cytosine) have the following meanings:

[0612] R: G or A

[0613] Y: T or C

[0614] M: A or C

[0615] K: G or T

[0616] S: G or C

[0617] W: A or T

[0618] H: A, C, or T

[0619] B: G or T or C

[0620] V: G, C, or A

[0621] D: G or A or T

[0622] N: G, A, T, or C

[0623] In SEQ ID NO: 1 through SEQ ID NO: 7, the SNP nucleotides of the resistance donor are shown in bold and underlined. SNP markers can be designed to the forward (+) strand or the reverse (-) strand of double-stranded DNA, which has been designated as forward or reverse relative to a reference genome sequence. For example, in the reference genome sequence of melon (found on melonomics.net, DHL92 version 4), SEQ ID NO: 4 is the reverse strand, and SNP_04 is “T” on the reverse strand and “A” on the forward strand (the forward strand is the complementary strand of SEQ ID NO: 4). SNP haplotypes or SNP donor nucleotides are preferably given relative to the forward strand in the reference genome, regardless of whether the sequence provided herein is the reverse strand.

[0624] SEQ ID NO 1: Sequence of a ToLCNDV-ES resistance donor plant containing SNP_01.

[0625] SEQ ID NO: 1 (forward strand in the reference genome):

[0626] TCAAAGGAGAGATTCTGATCTTCACTGCCAGGGCCAGCTTTCCTTTTTATTCGGTAGTATTTCTTTTGGTAATATTTGTA C GGGCTGACTCTTTTAGATGATGAAGCTGCTTTATTAGCCGCAAATTGTTCTATCCTTTCATTAATGCTGAGATTTTGTCT

[0627] SEQ ID NO 2: Sequence of a ToLCNDV-ES resistance donor plant containing SNP_02.

[0628] SEQ ID NO: 2 (Reverse strand in reference genome)

[0629] CATCCACCTTTACAACATTCCTGCACCATGAGATCAGTAAATAGTTATAAGTAATATATATTATATGAATGAAATTAAATATACATGAGCTAGAGATAATGGTGAAGTCATGAGAAAGAGATATGGTACGCACCGCAGAATTACCACAAGATATGAAATCAGCTAAGTTGTCGGGAAATCTAGCCAATGATTTGCATCCTGCGGCACTCGTACAAATTACACCTTCTGGAACCTTTGAAAT C TCTTCRAGCAACTCACAATCCATTGTATAAAGATATTTCAGGGATTTAAAATTAATAATACAYGAGGGTAKTCTACAAAAGTTGTTTTCGGACAAGTCCAACTCTTTCAAYGAAGGGGCAACATAAACAATTGTTTCTAAGAAATCCAAATTTGTTATCTTGCAACCTACAAGACGTAACTTGGTTAGGTAAAAAAGTGAGGAAGGTAAAGACGGATGAAGMCTCC

[0630] SEQ ID NO 3: Sequence of a ToLCNDV-ES resistance donor plant containing SNP_03.

[0631] SEQ ID NO 3 (forward strand in the reference genome)

[0632] TACAAGTGACAAAACCGAGTCGATCCGTAGTGAGCACGAAGATTTGTTTGTCAAAGTTAATTCTTTAGTTAATGAGATAA G TACCGAAGATGGTAAAAAAGAACGAAAACAGGGACCCACATCCTTTTGACTTGATTGCAGAGAGAAGATGAAAGAAATT

[0633] SEQ ID NO 4: Sequence of a ToLCNDV-ES resistance donor plant containing SNP_04.

[0634] SEQ ID NO 4 (reverse strand in the reference genome)

[0635] TCTCATTTTCAATGCCATGTTTTTTTTACTCATTTATCAACGAGCACACTGTACAATTTTTAAGATCCATTCTCTCTCAAGATGCCATTTAATTGTGTGT A CATCATTCTTTAATTTTGATATGATTATGTACTTGACAACAGGAGGGAGGCTGATCTTATTTGAGATCTTGTTAAAGAAGTGTTATCTACAATAAATCGC

[0636] SEQ ID NO 5: Sequence of a ToLCNDV-ES resistance donor plant containing SNP_05.

[0637] SEQ ID NO: 5 (Reverse strand in the reference genome)

[0638] CACTATAAACCTTATTCTTTTTAAGAAATCATGTTTCAAACCCAATAAAAGTAAGTTGATACATTGTCCTTGTAGAAATTGTCAAAACTAAATACAGTAAA T AAACAACTTTAAAATTTTGTATCATGTTATATTACAATATGGCCCATCTCACGTTTTCCCAATTGTCTTGAGAATTCAATGATATTTCATTCCAATATAA

[0639] SEQ ID NO 6: Sequence of a ToLCNDV-ES resistance donor plant containing SNP_06.

[0640] SEQ ID NO: 6 (Reverse strand in reference genome)

[0641] CCCATCTTCCCTTTGCCACTCTAAAATAATAGAGGTTATCTTTTTGTGTTTATTACATTTGAGTAAACTTATCAAATAAAATTGGGGAGTAGGAGATC A ACTGACGGAGAGAAATAAAGGGGAGAAGAAGGAAAATCCAGAAGTGAAAATGGAGGAAAAGGAATGGAAAAAAATTAGAGGGGTAAAGAATGAGAAAACAA

[0642] SEQ ID NO 7: Sequence of a ToLCNDV-ES resistance donor plant containing SNP_07.

[0643] SEQ ID NO: 7 (forward strand in the reference genome)

[0644] TTATCTTCTTGAACTGCCGAAGAAACTGCTTCTTTCTCCTTACCTTCATGTTCAGTCGAGAACTCTTCTACATTCCCCTC C GTTTTCTCTACATCAGACACACCTACTTCCTTCTCCTCACATTCAGGCAAAGCAGAATCTGCTTCCATATTTGAATCTTG

[0645] SEQ ID NO 8: TIR-NBS-LRR protein of the ToLCNDV-ES resistance donor of this application

[0646] MGSSALPVESSSSSSSSSPNFLYYYDYDVFFSFRGEDTRSSFISHLHMALRLKGVNVFID

[0647] DKLKRGDQISESLLKSIERSRLSLLVIFSKNYASSTWCLDELVKIIEYKKSKSQAVLPVFY

[0648] KVDPSEVRKQTGGFGEALAKHEANKLLTNKIQPWKEALTFAAGLSGWDLANCKDEAELIQ

[0649] EIVKRVLSVLNPMQLLHVAKHPVGIDFRLRKIEELVSHIGSEGVNMVGMYGIGGIGKTTL

[0650] AKALYNKIANQFEGCCFLQDVRREASKHGLVKLQETLLNDILKEDLKVVSRDRGINIIRS

[0651] RLCSKKVLIVLDDVDDREQLEALVGGRDWFGRGSKIIVTTRNDHLLSSHGFDKKHKIQEL

[0652] NQDHALELFSWHASKKSHPSSNYLCLSERATNYCKGLSLALVVLGSFLRGRDQAEWNCIL

[0653] DEFETSLRKDIKDVLQLSFDGLEDKAKDIFLDISCLLVGEEYNCAKKMLSACHLNIDFGI

[0654] MILMDLSLVIVETDRVQMHELIQQMGCSIVHNESSEPGKRSRLWLVQDIWEVFVNNSGTD

[0655] AVKAIKLDLPNPTKLNVDPQAFRSMKNLRLLIIRNAQFCRKIKYLPNSLKWIEWLGFAHR

[0656] SLPSCFITKNLVGLDLRHSSIKRFGKRLKGCERLKHVDLSYSTLLEQINDFSPASNLEEL

[0657] HLINCTNLGMIDKSVFSLHKLSVLNLDGCCNLQKLPRGYFMLSSLKELNLCYCKKLEKIP

[0658] DLSAASNLKRLYLQECTNLRVIHESVGSLDKLNHLDLRQCTKLVKLPSYLRLKSLSNLLL

[0659] SGCCKLESFPTIAENMKSLRELDMDFTAIKELPSSIGYLTNLSILKLNGCTNLISLPNTI

[0660] YLLRNLENLLLSGCSIFGMFPHTWDPTIPTIQQVCSPSNMMETASWSLEFPHLLVPNESL

[0661] CAHFTLLDLESCNISNAKFLELLCDVAPFLSDLRLSENKFSSLPSCLHKFMSLWNLELRN

[0662] CKFLQEIPNLPENIQKMDASGCESLARNPDNIVDIISKKQDLTLGEISREFLLTGIEIPE

[0663] WFSYKTTSNLVTASFRHYPDMERTLAACVSFKVNGDSSKRGAQISCSIFICSKLHSSFSR

[0664] PFLPSKSEYMWLVTTSLAWGSMEVNDWNKVLVWFEVHEAHGEVNVTITRYGVHVTEELHG

[0665] IQMDVKWPMVNYADFYQLEKLQSLDIEDLLLKSFLETVSCLSNSKAAMLHAGNYDPEAII

[0666] DSNIQPMIFPLHVTNNDGTYICGGMGGTALANSLCNKFKGMEGQCGEALDNSTSFFHIKR

[0667] RQLLSYSWSPAVHHRKCGDGERGTNITTHTISSKRYLILLREAKSYQDVHDWFYTHCWIK

[0668] ASYCSYDGRGDGVILIEGVDTSLL

[0669] SEQ ID NO 9: TIR-NBS-LRR protein of the ToLCNDV-ES resistance donor of US11591611 / NCIMB42585

[0670] MGSSALPVESSSSSSSSSPNFLYYYDYDVFFSFRGEDTRSSFISHLHMALRLKGVNVFID

[0671] DKLKRGDQISESLLKSIERSRLSLVIFSKNYASSTWCLDELVKIIECMKPKGQLVLSVFY

[0672] KVDPSVVWKQTGGFGEALAKHEATKLTTNKIKPWKEALTIAAGLSGWDLANCKDEAELIQ

[0673] EIVKRVLSVLNPMQLLHVAKHPVGIDFRLRKIEELVSHIGSEGVNMVGMYGIGGIGKTTL

[0674] AKALYNKIANQFEGSCFLQDVRREASKHGLVKLQETLLHDILKEDLKVVSRDRGINIIRS

[0675] RLCSKKVLIVLDDVDDREQLEALVGGRDWFGRGSKIIVTTRNHEHLLFSHGFDDQKHKIQE

[0676] LNQDHALELFSWHAFKKSHPSSNYLGLSERATNYCKGLSLALVVLGSFLRGRDQAEWNCI

[0677] LDEFETSLRKDIKDVLQLSFDGLEDKAKEIFLDISCLLVGEEINCAKKMLSACHLNIDFE

[0678] IMILMDLSLVTIETDRVQMHELIQQMGRSIVHNESSEPGKRSRLWLVQDIWEVFVNNSGT

[0679] DAVKAIKLDLPNPTKLNVDPQAFRSMKNLLRLIIRNAQFCRKIKYLPNSLKWIEWRGFAH

[0680] RSLPSCFITKNLVGLDLRHSSIKRFGKRLKGCERLKHVDLSYSPLLEKIPNLSAASNLEQ

[0681] LYLINCKNLGMIDKSVFSLDKLNVLNLDGCCNLQKLPRGYFMLSSLRYLNLSYKKKLEKI

[0682] PDLSAASNLKRLYLQECTNLRVIHESVGSLDKLDHLDLRQCTKLVKLPSYLRLKSLEYLS

[0683] LSGCCKLESFPTIAENMKSLWGLDLDFTAIKELPSSIGYLTKLSILKLNGCTNLISLPNT

[0684] IYLLRNLENLLLSGCSIFGMFPHTWDPTIPTIQQVCSPSKMMETASWSLEFPDLLVPNES

[0685] LCAHFTLLDLESCNISNAKFLELLCDVAPFLSDLRLSENKFSSLPSCLHKFMSLWNLELR

[0686] NCKFLQEIPNLPENIQKMDASGCESLARNPDNIVDIISKKQDLTLGEISREFLLTGIEIP

[0687] EWFSYKTTSNLVTASFRHYPDMERTLAACVSFKVNGDSSKRGAQMSCSIFICSKLHSSFS

[0688] RPFLPSKSEYMWLVTTSLAWGSMEVNDWNKVLVWFEVHEAHSEVNATITRCGVHVTEELH

[0689] GIQMDVKWPMVNYADFYQLEKLQSLDIEDLLLKSFLETVSCLSNSKAAMLHAGNYDPEAI

[0690] IDSNIQPMIFPLHVTYNGETVICGMEGMGDTTLANSLCNKFKGMEGQCGEALDNSTSFFH

[0691] IKRRQLLSYSWSPAVHHRKCGDGERGTNITTHTISSKRYLILLREAKSYQDVHDWFYTHC

[0692] WIKASYCSYDGRGDGVFLIKGVDTSLL

[0693] SEQ ID NO 10: TIR-NBS-LRR protein of reference genome DHL92

[0694] MGSSALPVESSSSSSSPNFLYYYDYDVFFSFRGEDTRSSFISHLHMALRLKGVNVFID

[0695] DKLKRGDQISESLLKSIERSRLSLVIFSKNYASSTWCLDELVKIIEYKKSKSQAVLPVFY

[0696] KVDPSEVRKQTGGFGEALAKHEANKLLTNKIQPWKEALTFAAGLSGWDLANCKDEAELIQ

[0697] EIVKRVLSVLNPMQLLHVAKHPVGIDFRLRKIEELVSHIGSEGVNMVGMYGIGGIGKTTL

[0698] AKALYNKIANQFEGCCFLQDVRREASKHGLVKLQETLLNDILKEDLKVVSRDRGINIIRS

[0699] RLCSKKVLIVLDDVDDREQLEALVGGRDWFGRGSKIIVTTRNEHLLFSHGFDDQKHKIQE

[0700] LNQDHALELFSWHAFKKSHPSSNYLGLSERATNYCKGLSLALVVLGSFLRGRDQAEWNCI

[0701] LDEFETSLRKDIKDVLQLSFDGLEDKAKDIFLDISCLLVGEEYNCAKKMLSACHLNIDFG

[0702] IMILVDLSLVTIETDRVQMHELIQQMGRSIVHNESSEPGKRSRLWLVQDIWEVFVNNSGT

[0703] DAVKAIKLDLPNPTKLNVDPQAFRSMKNLRLLIIRNAQFCRKIKYLPNSLKWIEWRGFAH

[0704] RSLPSCFITKNLVGLDLRHSSIKRFGKRLEGCERLKHVDLSYSTLLEQINDFSPASNLEE

[0705] LHLINCTNLGMIDKSVFSLYKLSVLNLDGCCNLQKLPRGYFMLSSLKELNLCYCKKLEKI

[0706] PDLSAASNLKRLYLQECTNLRVIHESVGSLDKLNHLDLRQCTKLVKLPSYLRLKSLSNLL

[0707] LSGCCKLESFPTIAENMKSLRELDMDFTAIKELPSSIGYLTNLSILKLNGCTNLISLPNT

[0708] IYLLRNLENLLLSGCSIFGMFPHTWDPTIPTIQQVCSPSKMMETASWSLEFPHLLVPNES

[0709] LCAHFTLLDLESCNISNAKFLELLCDVAPFLSDLRLSENKFSSLPSCLHKFMSLWNLELR

[0710] NCKFLQEIPNLPENIQKMDASGCESLARNPDNIVDIISKKQDLTLGEISREFLLTGIEIP

[0711] EWFSYKTTSNLVTASFRHYPDMERTLAACVSFKVNGDSSKRGAQISCSIFICSKLHSSFS

[0712] RPFLPSKSEYMWLVTTSLAWGSMEVNDWNKVLVWFEVHEAHSEVNATITRCGVHVTEELH

[0713] GIQMDVKWPMVNYADFYQLEKLQSLDIEDLLLKSFLETVSCLSNSKAAMLHAGNYDPEAI

[0714] IDSNIQPMIFPLHVTNNDGTYICGGMGGTALANSLCNKFKGMEGQCGEALDNSTSFFHIK

[0715] RRQLLSYSWSPAVHHRKCGDGERGTNITTHTISSKRYLILLREAKSYQDVHDWFYTHCWI

[0716] KASYCSYDGRGDGVILIEGVDTSLL

[0717] SEQ ID NO 11: Genomic DNA encoding the TIR-NBS-LRR protein of SEQ ID NO: 8 that is the ToLCNDV-ES resistance donor of the present invention

[0718] ATGGGTTCTTCTGCTCTTCCAGTTGAATCTTCTTCT

[0719] TCTTCTTCTTCTTCTTCTCCCAACTTTCTCTATTACTACGATTATGATGTGTTTTTTTAGT

[0720] TTCAGGAGGAGAAGACACTCGCTCCAGTTCATCAGTCATCTTCATATGGCCTTGCGTCTA

[0721] AAGGGAGTCAACGTCTTCATAGACGACAAACTCAAAAGGGGTGACCAAATCTCTGAGTCT

[0722] CTTTCTAAATCTATAGAGCGATCTAGACTTTCACTCGTTATTTTCTCTAAAAATTATGCA

[0723] TCTTCAACTTGGTGTTTGGATGAACTGGTGAAAATAATTGAGTATAAAAAATCCAAAAGT

[0724] CAAGCGGTTTTGCCGGTGTTCTACAAGGTGGATCCGTCCGAGGTTCGAAAACAAACCGGT

[0725] GGGTTTGGGGAAGCATTAGCCAAACATGAAGCTAATAAGTTATTGACCAACAAGATTCAA

[0726] CCATGGAAGGAAGCTTTGACTTTTGCTGCTGGTTTGTCTGGTTGGGATCTAGCAAATTGG

[0727] TATTTCTTTTTTTAATTTCCCAAGACTCATTGTCTAAGTGAAGTTTAAATTTATCTACTA

[0728] TTTCCTATTTTCATTTTTTTTTTTTGTATGATGTTACTTTACAACAGCAAGGATGAGGCT

[0729] GAACTTATCCAAGAAAATTGTTAAACGAGTGTTGTCTGTATTAAATCCAATGCAATTACTA

[0730] CATGTAGCCAAGCATCCAGTTGGAATTGATTTTCGATTAAGGAAAATTGAGGAGTTGGTC

[0731] TCTCATATTGGGTCCGAGGGTGTTAACATGGTGGGGATGTATGGCATTGGAGGCATTGGT

[0732] AAGACCACTTTAGCTAAGGCTTTGTACAACAAAATTGCTAACCAATTTGAAGGATGTTGC

[0733] TTTCTACAAGATGTTAGACGAGAAGCTTCAAAGCATGGGCTCGTTAAACTACAAGAAACC

[0734] TTACTCAATGACATCTTAAAGGAGGATTTGAAGGTTGTCAGCCGTGATAGAGGAATTAAC

[0735] ATCATAAGGAGTAGACTGTGTTCAAAGAAAGTTCTTATAGTTCTTGATGATGTGGATGAT

[0736] CGTGAGCAATTAGAAGCACTGGTTGGTGGTCGTGATTGGTTTGGTCGAGGTAGCAAAATC

[0737] ATTGTGACGACAAGGAATGATCATTTACTTTCTAGCCATGGATTTGATAAAGGCATAAA

[0738] ATTCAAGAATTGAATCAAGATCATGCTCTTGAGCTTTTTAGTTGGCACGCTTCTAAGAAA

[0739] AGCCATCCATCAAGTAATTATCTATGCCTTTCAGAACGTGCTACAAATTATTGTAAAGGT

[0740] CTATCTTTGGCACTCGTTGTTTTGGGTTCTTTCCTTCGTGGCAGAGATCAAGCAGAATGG

[0741] AACTGTATATTAGATGAATTTGAAACCTCTTTGAGAAAAGATATTAAAGATGTTCTTCAA

[0742] TTAAGTTTTGATGGACTTGAAGACAAAGCAAAGGATATTTTCCTTGATATTTCTTGTTTA

[0743] CTTGTGGGAGAAGAATACAATTGTGCTAAAAAAATGTTGAGTGCATGCCATTTGAACATA

[0744] GATTTTGGAATTATGATACTCATGGATCTTTCTCTTGTTATTGTTGAAACGGATAGAGTG

[0745] CAAATGCATGAGTTAATACAACAAATGGGTTGTAGCATAGTTCATAATGAATCATCTGAG

[0746] CCTGGAAAGAGGAGTAGGTTGTGGTTGGTGCAGGACATTTGGGAGGTGTTTGTTAATAAT

[0747] TCAGTGAGTAACTCCTACCTAAAGTATTTAATAATTTGGACTTCACCCTGATAATATATAA

[0748] ATGTTTGTCAAAGTAATTAAATTGTCAGTAATGTTAAATTACTAAATATATGACTTTGTA

[0749] GGGAACAGATGCAGTTAAAGCCATAAAGTTGGACTTGCCTAATCCCACAAAGCTAAATGT

[0750] AGATCCACAAGCATTTAGAAGTATGAAAAATTTGAGATTGCTTATCATTCGAAATGCACA

[0751] ATTTTTAGAAAGATTAAGTACCTACCTAATAGCTTAAAGTGGATTGAGTGGCTTGGATT

[0752] TGCTCATCGATCTTTGCCGTCATGCTTCATTACCAAAAATCTTGTTGGACTTGATTTGCG

[0753] ACATAGCTCCATCAAAAGATTTGGGAAAAGGCTCAAGGTAAAATATTTCTTTTTGCAT

[0754] CTGTATTTGACTGGAGATTTCTTCATTGATTTTCTTAAAAAGTTTATGGGTAGCTTAA

[0755] CTGCACCGGAAAAAAAAAATCACAATAGGTGATTTAGCAAAATTTAACATGGATTAACC

[0756] CGTGCTAATTATTATTTTAAAGAATTTGCAAATTAATTATCATTAATTATTTAATATATATTAAT

[0757] GTGATCTATCGCACAACCATAGACTTATGCTAGTCATGATCTATCATTAATATACTCC

[0758] TATTAGTGATATGATCTACATGGATTAATTCCAAGGGGTCTTTTGGGGAAAGGGTTGGAT

[0759] TATGGGAGTTAAGGTTATGATAAAACTAGTGTTATGACAATCTAGGATTACGATAGA

[0760] TGTATTTAGTGGAAGGTTATGGAGGAAGTATTATGATAAAATATGTTTGGGAGAAGGGT

[0761] TATGTGGGTAGGGTTATGATATATATATATATATATATATTATTATTATTCAT

[0762] ATTATATCAAATCAACATTTCGTATTATTTACTATTGTCCTAGTTTTCAT

[0763] AAAATAATTTGCCTTAATTTATTTAACTACGTTATTCAATTTCAACATACACATTGTTTA

[0764] TTTTCAACATTTTTTAAGTATACGTTACGTTTCAATAATTTGGTTTTCATGAATATGACA

[0765] CACAACCTGTAAATATGTAAATAAGGGTAAATAACAATTGAAAATGCGAATTACCGAAAA

[0766] CAGAAAATAATAATCAACGCTTTTACAAACTCTAGTTTTGTTTTCTAGTGAAAAATTACA

[0767] ATGAATCTAGTTTTAGGTTTCTCTTCAATTTATTTTAGTCTACTTTTAAAATATTAACTT

[0768] TATTTTTTATTCTTTTTAACGGTGATTTATACTTTCAAAATATTTATTTTAGTCATTTTG

[0769] CAACAATTATTGGGTAAAAAATTCATGAATCTACCGCCTCTATAACAAAAAAATTTGTTA

[0770] TATTAATTTAACCAAGCTTAATTTAATAACCAAATTAACATTATGAAAGTGGGAAAATAT

[0771] AGACAAACTGCATCTAAATAATAGATTTGTTAAATTATAGCCCCACCAATTTTTAAATTA

[0772] TTCAAATAGTCCTTATGATTTTCTAATTGTTTTAATCAATTTATGGTAGTTTGTTAAAAT

[0773] TTTACCATTTTTTTTATTTAATTGATATTCCTTAGATATTTATCCTAGCTAACTAAAAAA

[0774] AAAACAATGAGATTGAGATTTATATTAAAATTTTAATTTCATAAATATATTGATATGAATT

[0775] CAAACATAGAGTTGATTGAATGTAACTAAATTAAAAATTTAATTGTGAAATCACTATAAA

[0776] TTATCTAATTCAAGAGTAAAGGAAAAAATTTAAATTTTAATAATGTAATTATACCAAATT

[0777] CATAATTAAAAAACAAGCTGTGTTTTTAAAATAAAAGATTAAAATGAACAAAACTCATA

[0778] AATATAAAAAATTAAATGAGAAAAAATGAATTGAGGAAGGGTTGAAGAAAGGTTGAGG

[0779] GGATAAAACTAAGGTTATGATAACCCTATAATTAAAAAGACTAAACTAACATTCTCATTCT

[0780] GTTTAAGGTATCAAATTAAATGGTTAGTTTACAATCTACTAAATTCAAGATAATATATT

[0781] TAGGTATAATGGTTACTTTACAATAATATTAATTAATTAATTACTTATAAACTAAAATTT

[0782] AATTCAAAACTATGTTTAATTTAACTAATTCTCTCGTGCCTAAATTATATTTTTTTAAAC

[0783] TTTTGTATTAAGTTGCATTTTAAACTTATAAAACACACTTACTATCGATATGTATCAAAC

[0784] TAATAGTTGTAAAAAGTGCATTGTTGCTAAACATGAGTACAGCTGAACTGACATAATATTG

[0785] ATTAATGGTTATTATACAATTTCTTTTACCTATATTCTATATTTTTCTATATTTACGATT

[0786] GTTGTTTTAGCCACTATTTTTATCTAAATTTTGTTTTGTATGGCAATTACATACCTCCTTT

[0787] TCCTTTTGGTTTTTTTTTTTCAATGTGTTTTAGGGTTGTGAAAGGTTGAAGCATGTTGAT

[0788] CTTAGCTACTCTACTTTATTAGAGCAAATTAATGATTTCTCTCCGGCATCAAATCTTGAA

[0789] GAATTGCATCTCATCAATTGCACAAATTTAGGAATGATAGATAAGTCTGTTTTTTTCTCTC

[0790] CATAAGCTTAGTGTCCTAAACCTTGATGGTTGTTGTAACCTTCAAAAGCTTCCAAGAGGC

[0791] TATTTCATGTTAAGTTCTCTTAAAGAATTGAATCTCTGTTACTGCAAAAAGCTTGAAAAA

[0792] ATTCCAGACTTATCTGCAGCATCAAACCTTAAGAGATTGTATCTCCAAGAATGCACAAAT

[0793] TTAAGAGTGATTCATGAATCTGTTGGATCTTTGGATAAGCTTAATCATCTGGACCTTAGA

[0794] CAATGCACTAAACTGGTAAAGCTTCCAAGCTATCTCAGGTTAAAGTCTCTTTCCAATTTA

[0795] TTACTTTCTGGGTGTTGTAAGCTTGAAAGCTTCCCAACAATTGCTGAAAACATGAAATCT

[0796] TTAAGGGAATTGGATATGGATTTTACTGCCATAAAGGAGTTACCTTCATCAATTGGATAT

[0797] CTTACTAACCTTTCTATATTAAAACTTAACGGTTGCACAAACCTCATCTCCCCTTCCCAAT

[0798] ACAATTTATTTGTTAAGGAATCTTGAGAATCTTCTTCTTAGTGGCTGTTCTATATTTGGA

[0799] ATGTTTCCCCATACATGGGACCCAACCATCCCAACCATCCAACAAGTATGCTCTCCTTCA

[0800] AATATGATGGAAACAGCTTCCTGGAGCTTAGAATTTCCCCATTTACTAGTACCAAATGAA

[0801] AGTTTATGTGCACATTTCACTTTGTTGGATCTTGAATCTTGCAACATATCAAATGCAAAA

[0802] TTTTTAGAATTATTATGTGATGTTGCCCCTTTCTTATCTGATCTACGCTTGTCTGAAAAC

[0803] AAATTCTCTAGTTTACCCTCATGTCTCCACAAGTTCATGTCCTTGTGGAATCTTGAATTA

[0804] AGGAATTGCAAGTTTCTTCAAGAAAATTCCAAACCTTCCTGAGAATATACAAAAAATGGAT

[0805] GCCAGTGGTTGTGAATCGTTGGCTCGAAATCCAGATAACATTGTGGATATAATATCAAAA

[0806] AAACAGGTTCGCCTCTAATTTCCATTCAATTTATATTCTTATCTTGTAAACAATTTAATG

[0807] CATTATGAATTCTTGTTCTCTATAGGACCTCACATTGGGTGAGATTTCAAGAGAGTTTTT

[0808] ATTAACGGGGATTGAGATTCCAGAATGGTCAGCTATAAGACTACATCCAATTTAGTGAC

[0809] AGCTAGCTTTCGTCACTATCCAGACATGGAAAGAACTTTGGCTGCCTGTGTTAGTTTCAA

[0810] AGTGAATGGAGATTCATCTAAAAGAGGAGCCCAAATTTCATGTAGTATATTCATCTGCAG

[0811] TAAACTCCATTCTTCATTTTCAAGACCATTTCTTCCATCAAAATCAGAATATATGTGGTT

[0812] AGTAACAACTTCTCTAGCGTGGGGTTCCATGGAGGTGAATGATTGGAATAAAGTTTTAGT

[0813] ATGGTTTGAGGTTCATGAAGCACATGGTGAGGTTAATGTAACTATAACAAGGTATGGTGT

[0814] CCATGTCACTGAAGAGCTCCATGGGATACAAATGGATGTCAAGTGGCCGATGGTAAATTA

[0815] TGCTGATTTTTATCAACTGGAGAAAATTGCAAAGTCTGTAAGTTGATTGTTAACTTGTTAT

[0816] TTATTTATTCTCTTTCTCTTTTTCTTTTTTTTTTTTTTTTTTTTTTTTTGGAGTGAAAGTA

[0817] TATAATAGATCTCAAAGGGGGAGATGAGTATGTTATGATTTTGTTTGTTTAATGGCATAG

[0818] GGATATTGAGGATCTTCTTCTCAAAAGCTTTTTAGAAACAGTCTCTTGCCTGTCAAATTC

[0819] CAAAGCAGCAATGTTACATGCAGGAAATTATGATCCAGAAGCAATAATTGATTCCAACAT

[0820] ACAACCTATGATATTTCCATTGCACGTAACAAATAATGATGGCACATATATATGTGGAGG

[0821] CATGGGAGGCACTGCACTTGCCAACTCTTTATGCAATAAATTTAAAGGGATGGAGGGCCA

[0822] ATGCGGTGAAGCTTTAGATAATTCTACAAGCTTTTTCCATATCAAAAGAAGACAACTCCT

[0823] GAGCTATTCCTGGTCGCCGGCGGTCCACCATCGTAAGTGTGGAGATGGTGAAAGAGGAAC

[0824] GAATATCACAACCCACACAATATCCTCCAAACGCTATTTGATACTCCTTCGTGAAGCCAA

[0825] GAGCTATCAGGATGTACATGACTGGTTTTATACACATTGTTGGATAAAAGCTTCATATTG

[0826] CAGTTATGACGGAAGAGGTGATGGTGTGATTCTGATTGAAGGGGTTGATACATCCTTGCT

[0827] CTGA

[0828] SEQ ID NO 12: cDNA (mRNA) of SEQ ID NO: 8

[0829] ATGGGTTCTTCTGCTCTTCCAGTTGAATCTTCTTCT

[0830] TCTTCTTCTTCTTCTTCTCCCAACTTTCTCTATTACTACGATTATGATGTGTTTTTTAGT

[0831] TTCAGGAGGAGAAGACACTCGCTCCAGTTCATCAGTCATCTTCATATGGCCTTGCGTCTA

[0832] AAGGGAGTCAACGTCTTCATAGACGACAAACTCAAAAGGGGTGACCAAATCTCTGAGTCT

[0833] CTTTCTAAATCTATAGAGCGATCTAGACTTTCACTCGTTATTTTCTCTAAAAATTATGCA

[0834] TCTTCAACTTGGTGTTTGGATGAACTGGTGAAAATAATTGAGTATAAAAAATCCAAAAGT

[0835] CAAGCGGTTTTGCCGGTGTTCTACAAGGTGGATCCGTCCGAGGTTCGAAAACAAACCGGT

[0836] GGGTTTGGGGAAGCATTAGCCAAACATGAAGCTAATAAGTTATTGACCAACAAGATTCAA

[0837] CCATGGAAGGAAGCTTTGACTTTTGCTGCTGGTTTGTCTGGTTGGGATCTAGCAAATTGC

[0838] AAGGATGAGGCTGAACTTATCCAAGAAATTGTTAAACGAGTGTTGTCTGTATTAAATCCA

[0839] ATGCAATTACTACATGTAGCCAAGCATCCAGTTGGAATTGATTTTCGATTAAGGAAAATT

[0840] GAGGAGTTGGTCTCTCATATTGGGTCCGAGGGTGTTAACATGGTGGGGATGTATGGCATT

[0841] GGAGGCATTGGTAAGACCACTTTAGCTAAGGCTTTGTACAACAAAATTGCTAACCAATTT

[0842] GAAGGATGTTGCTTTCTACAAGATGTTAGACGAGAAGCTTCAAAGCATGGGCTCGTTAAA

[0843] CTACAAGAAACCTTACTCAATGACATCTTAAAGGAGGATTTGAAGGTTGTCAGCCGTGAT

[0844] AGAGGAATTAACATCATAAGGAGTAGACTGTGTTCAAAGAAAGTTCTTATAGTTCTTGAT

[0845] GATGTGGATGATCGTGAGCAATTAGAAGCACTGGTTGGTGGTCGTGATTGGTTTGGTCGA

[0846] GGTAGCAAAATCATTGTGACGACAAGGAATGATCATTTACTTTCTAGCCATGGATTTGAT

[0847] AAAAAGCATAAAATTCAAGAATTGAATCAAGATCATGCTCTTGAGCTTTTTAGTTGGCAC

[0848] GCTTCTAAGAAAAGCCATCCATCAAGTAATTATCTATGCCTTTCAGAACGTGCTACAAAT

[0849] TATTGTAAAGGTCTATCTTTGGCACTCGTTGTTTTGGGTTCTTTCCTTCGTGGCAGAGAT

[0850] CAAGCAGAATGGAACTGTATATTAGATGAATTTGAAACCTCTTTGAGAAAAGATATTAAA

[0851] GATGTTCTTCAATTAAGTTTTGATGGACTTGAAGACAAAGCAAAGGATATTTTCCTTGAT

[0852] ATTTCTTGTTTACTTGTGGGAGAAGAATACAATTGTGCTAAAAAAATGTTGAGTGCATGC

[0853] CATTTGAACATAGATTTTGGAATTATGATACTCATGGATCTTTCTCTTGTTATTGTTGAA

[0854] ACGGATAGAGTGCAAATGCATGAGTTAATACAACAAATGGGTTGTAGCATAGTTCATAAT

[0855] GAATCATCTGAGCCTGGAAAGAGGAGTAGGTTGTGGTTGGTGCAGGACATTTGGGAGGTG

[0856] TTTGTTAATAATTCAGGAACAGATGCAGTTAAAGCCATAAAGTTGGACTTGCCTAATCCC

[0857] ACAAAGCTAAATGTAGATCCACAAGCATTTAGAAGTATGAAAAATTTGAGATTGCTTATC

[0858] ATTCGAAATGCACAATTTTGTAGAAAGATTAAGTACCTACCTAATAGCTTAAAGTGGATT

[0859] GAGTGGCTTGGATTTGCTCATCGATCTTTGCCGTCATGCTTCATTACCAAAAATCTTGTT

[0860] GGACTTGATTTGCGACATAGCTCCATCAAGATTTGGGAAAAGGCTCAAGGGTTGTGAA

[0861] AGGTTGAAGCATGTTGATCTTAGCTACTCTACTTTATTAGAGCAAATTAATGATTTCTCT

[0862] CCGGCATCAAATCTTGAAGAATTGCATCTCATCAATTGCACAAATTTAGGAATGATAGAT

[0863] AAGTCTGTTTTTTTCCTCCATAAGCTTAGTGCTCTAAAACCTTGATGGTTGTTGTAACCTT

[0864] CAAAAGCTTCCAAGAGGCTATTTCATGTTAAGTTCTCTTAAAGAATTGAATCTCTGTTAC

[0865] TGCAAAAAGCTTGAAAAAATTCCAGACTTATCTGCAGCATCAAACCTTAAGAGATTGTAT

[0866] CTCCAAGAATGCACAAATTTAAGAGTGATTCATGAATCTGTTGGATCTTTGGATAAGCTT

[0867] AATCATCTGGACCTTAGACAATGCACTAAACTGGTAAAGCTTCCAAGCTATCTCAGGTTA

[0868] AAGTCTCTTTCCAATTTATTACTTTCTGGGTGTTGTAAGCTTGAAAGCTTCCCAACAATT

[0869] GCTGAAAACATGAAATCTTTAAGGGAATTGGATATGGATTTTACTGCCATAAAGGAGTTA

[0870] CCTTCATCAATTGGATATCTTACTAACCTTTCTATATTAAAACTTAACGGTTGCACAAAC

[0871] CTCATCTCCCTTCCCAATACAATTTATTTGTTAAGGAATCTTGAGAATCTTCTTCTTAGT

[0872] GGCTTGTTCTATATTTGGAATGTTTCCCCATACATGGGACCCAACCATCCCAACCATCCAA

[0873] CAAGTATGCTCTCCTTCAAATATGATGGAAACAGCTTCCTGGAGCTTAGAATTTCCCCAT

[0874] TTACTAGTACCAAATGAAAGTTTATGTGCACATTTCACTTTGTTGGATCTTGAATCTTGC

[0875] AACATATCAAATGCAAAATTTTTAGAATTATTATGTGATGTTGCCCCTTTCTTATCTGAT

[0876] CTACGCTTGTCTGAAAACAAATTCTCTAGTTTACCCTCATGTCTCCACAAGTTCATGTCC

[0877] TTGTGGAATCTTGAATTAAGGAATTGCAAGTTTCTTCAAGAAATTCCAAACCTTCCTGAG

[0878] AATATACAAAAAATGGATGCCAGTGGTTGTGAATCGTTGGCTCGAAATCCAGATAACATT

[0879] GTGGATATAATATCAAAAACAGGACCTCACATTGGGTGAGATTTCAAGAGAGTTTTTA

[0880] TTAACGGGGATTGAGATTCCAGAATGGTCAGCTATAAGACTACATCCAATTTAGTGACA

[0881] GCTAGCTTTCGTCACTATCCAGACATGGAAAGAACTTTGGCTGCCTGTGTTAGTTTCAAA

[0882] GTGAATGGAGATTCATCTAAAAGAGGAGCCCAAATTTCATGTAGTATATTCATCTGCAGT

[0883] AAACTCCATTCTTCATTTTCAAGACCATTTCTTCCATCAAAATCAGAATATATGTGGTTA

[0884] GTAACAACTTCTCTAGCGTGGGGTTCCATGGAGGTGAATGATTGGAATAAAGTTTTAGTA

[0885] TGGTTTGAGGTTCATGAAGCACATGGTGAGGTTAATGTAACTATAACAAGGTATGGTGTC

[0886] CATGTCACTGAAGAGCTCCATGGGATACAAATGGATGTCAAGTGGCCGATGGTAAATTAT

[0887] GCTGATTTTTATCAACTGGAGAAATTGCAAAGTCTGGATATTGAGGATCTTCTTCTCAAA

[0888] AGCTTTTTAGAAACAGTCTCTTGCCTGTCAAATTCCAAAGCAGCAATGTTACATGCAGGA

[0889] AATTATGATCCAGAAGCAATAATTGATTCCAACATACAACCTATGATATTTCCATTGCAC

[0890] GTAACAAATAATGATGGCACATATATATGTGGAGGCATGGGAGGCACTGCACTTGCCAAC

[0891] TCTTTATGCAATAAATTTAAAGGGATGGAGGGCCAATGCGGTGAAGCTTTAGATAATTCT

[0892] ACAAGCTTTTTCCATATCAAAAGAAGACAACTCCTGAGCTATTCCTGGTCGCCGGCGGTC

[0893] CACCATCGTAAGTGTGGAGATGGTGAAAGAGGAACGAATATCACAACCCACACAATATCC

[0894] TCCAAACGCTATTTGATACTCCTTCGTGAAGCCAAGAGCTATCAGGATGTACATGACTGG

[0895] TTTTATACACATTGTTGGATAAAAGCTTCATATTGCAGTTATGACGGAAGAGGTGATGGT

[0896] GTGATTCTGATTGAAGGGGTTGATACATCCTTGCTC

[0897] SEQ ID NO 13: Genomic DNA encoding SEQ ID NO: 9

[0898] ATGGGTTCTTCTGCTCTTCCAGTTGAATCTTCTTCT

[0899] TCTTCTTCTTCTTCTTCTCCCAACTTTCTCTATTACTACGATTATGATGTGTTTTTTTAGT

[0900] TTCAGGAGGAGAAGACACTCGCTCCAGTTCATCAGTCATCTTCATATGGCCTTGCGTCTA

[0901] AAGGGAGTCAACGTCTTCATAGACGACAAACTCAAAAGGGGTGACCAAATCTCTGAGTCT

[0902] CTTTCTAAATCTATAGAGCGATCTAGACTTTCACTCGTTATTTTCTCTAAAAATTATGCA

[0903] TCTTCAACTTGGTGTTTGGATGAACTGGTGAAAATAATTGAGTGTATGAAACCCAAAGGA

[0904] CAGCTAGTTTTGTCTGTCTTCTACAAGGTGGATCCGTCCGTGGTTTGGAAACAAACAGGT

[0905] GGGTTTGGTGAAGCATTGGCCAAACATGAGGCTACTAAGTTAACAACCAACAAGATTAAA

[0906] CCATGGAAGGAAGCTTTGACTATTGCTGCTGGTTTGTCTGGTTGGGATCTAGCAAATTGG

[0907] TATTTCTTTTTTTAATTTCCCAAGACTCATTGTCTAAGTGAAGTTTAAATTTATCTACTA

[0908] TTTCTTATTTTCATTTTTTTTTTTGTATGATGTTACTTTACAACAGCAAGGATGAGGCTG

[0909] AACTTATCCAAGAAAATTGTTAAACGAGTGTTGTCTGTATTAAATCCAATGCAATTACTAC

[0910] ATGTAGCCAAGCATCCAGTTGGAATTGATTTTCGATTAAGGAAAATTGAGGAGTTGGTCT

[0911] CTCATATTGGGTCCGAGGGTGTTAACATGGTGGGGATGTATGGCATTGGAGGCATTGGTA

[0912] AGACCACTTTGGCTAAGGCTTTGTACAACAAAATTGCTAACCAATTTGAAGGATCTTGCT

[0913] TTCTACAAGATGTTAGACGAGAAGCTTCAAAGCATGGGCTCGTTAAACTACAAGAAACCT

[0914] TACTCCATGACATCTTAAAGGAGGATTTGAAGGTTGTCAGCCGTGATAGAGGAATTAACA

[0915] TCATAAGGAGTAGACTGTGTTCAAAGAAAGTTCTTATAGTTCTTGATGATGTGGATGATC

[0916] GTGAGCAATTAGAAGCACTGGTTGGTGGTCGTGATTGGTTTGGTCGAGGTAGCAAAATCA

[0917] TTGTGACGACAAGGAATGAGCATTTACTTTTTAGCCATGGATTTGATGATCAAAAGCATA

[0918] AAATTCAAGAATTGAATCAAGATCATGCTCTTGAGCTTTTTAGTTGGCACGCTTTTAAGA

[0919] AAAGCCATCCATCAAGTAATTATCTAGGCCTTTCAGAACGTGCTACAAATTATTGTAAAG

[0920] GTTCTATCTTTGGCACTCGTTGTTTTGGGTTCTTTCCTTCGTGGCAGAGATCAAGCAGAAT

[0921] GGAACTGTATATTAGATTTGAAACCTCTTTGAAAAGATATTAAAGATGTTCTTC

[0922] AATTAAGTTTTGATGGACTTGAAGACAAAGCAAAGGAAATTTTCCTTGATATTTCTTGTT

[0923] TACTCGTGGGAGAAGAAATCAATTGTGCTAAAAAGATGTTGAGTGCATGCCATTTGAACA

[0924] TAGATTTGAAATTATGATACTCATGGATCTTTCACTTGTTACTATTTGAAACGGATAGAG

[0925] TGCAAATGCATGAGTTAATACAACAAATGGGTCGTAGCATAGTTCATAATGAATCATCTG

[0926] AGCCTGGAAAGAGGAGTAGTTGTGGTTGGTGCAGGACATTTGGGAGGTGTTTGTTAATA

[0927] ATTCAGTGAGTAACTCCTACCTAAAGTATTTAATTTGGACTTCACCTGATAATATAT

[0928] AAATGTTTGTCAAAGTAATTAAATTGTCAGTAATGTTAATTACTAAATATATGACTTTG

[0929] TAGGGAACAGATGCAGTTAAAGCCATAAAGTTGGACTTGCCTAATCCCACAAAGCTAAAT

[0930] GTAGATCCACAAGCATTTAGAAGTATGAAAATTTGAGATTGCTTATCATTCGAAATGCA

[0931] CAATTTGTAGAAAGATTAAGTACCTACCTAATAGCTTAAAGTGGATTGAGTGGCGTGGA

[0932] TTTGCTCATCGATCTTTGCCGTCATGCTTCATTACCAAAAATCTTGTTGGACTTGATTTG

[0933] CGACATAGCTCCATCAAAAGATTTGGGAAAAGGCTCAAGGTAAAATATATTTCTTTTTTGC

[0934] ATCTGTATTTGACTGGAGATTTCTTCATTGATTTTCTTAAAAAGTTTTAATGGGTAGCTT

[0935] AACTGCACCGGAAAAAAAAAATCACAAATAGGTGATTTAGCAAAATTTAACATGGATTAA

[0936] CCCGTGCTAATCACATATTTTTAAAGAATTTGCAAATTATATCAAAGTTTATTAATGATA

[0937] TAGTGATCTATCGCACAACCATAGACTTAGTGCTAGTCATATGATCTATCATTAATATACT

[0938] CCTATTAGTGATATGATCTACATGGATTAATTCCAAGGGGTCTTTTGGGGAAAGGGTTGG

[0939] ATTATGGAGTTAAGGTTATGATAAAACTAGTGTTATGACAAATCTAGGATTACGATAAA

[0940] GATGTATTTAGTGGAAGGGTTATGGAGGAAGTATTATGATAAAATATGTTTGGGAGAAGG

[0941] GTTATGTGGGTAGGGTTATGATAAATATATATATATATTAATTCATATTATAAATCAAAT

[0942] ACACAAAATTTCGTATATTTAATTTACTAAATTGTCCTAGTTTTCATAAAAATAATTTGCCT

[0943] TAATTTATTTAACTACGTTATTCAATTTCAACATACACATTGTTTATTTTCAACATTTTT

[0944] TAAGTATACGTTACGTTTCAATAATTTGGTTTTCATGAATATGACACACAACCTGTAAAT

[0945] ATGTAAATAAGGGTAAATAACAATTGAAAATGCGAATTACCGAAAACAGAAAATAATAAT

[0946] CAACGCTTTTACAAACTCTAGTTTTGTTTTCTAGTGAAAAATTACAATGAATCTAGTTTT

[0947] AGGTTTCTCTTCAATTTATTTTAGTCTACTTTTAAAATATTAACTTTATTTTTTATTCTT

[0948] TTTAACGGTGATTTATACTTTCAAAATATTTATTTTAGTCATTTTGCAACAATTATTGGG

[0949] TAAAAAATTCATGAATCTACCGCCTCTATAACAAAAAAATTTGTTATATTAATTTAACCA

[0950] AGCTTAATTTAATAACCAAATTAACATTATGAAAGTGGGAAAATATAGACAAACTGCATC

[0951] TAAATAATAGATTTGTTAAATTATAGCCCCACCAATTTTTAAATTATTCAAATAGTCCTT

[0952] ATGATTTTCTAATTGTTTTAATCAATTTATGGTAGTTTGTTAAAATTTTACCATTTTTTT

[0953] TATTTAATTGATATTCCTTAGATATTTATCCTAGCTAACTAAAAAAAAACAATGAGATTG

[0954] AGATTTATATTAAAATTTTAATTTCATAAATATATTGATATGAATTCAAACATAGAGTTG

[0955] ATTGAATGTAACTAAATTAAAAATTTAATTGTGAAATCACTATAAATTATCTAATTCAAG

[0956] AGTAAAGGAAAAAATTTAAATTTTAATAATGTAATTATACCAAATTCATAATTAAAAAAC

[0957] AAGCTGTGTTTTTAAAATAAAGATTAAAATGAACAAAACTCATAAATATAAAAATTAT

[0958] AAATGAGAAAAAAATGAATTGAGGAAGGGTTGAAGAAAGGTTGAGGGGATAAAACTAAGG

[0959] TTATGATAACCCTATAATTAAAGACTAAACTACATTCTCATTCTGTTTAAGGTATCAA

[0960] ATTAAATGTTTAGTTTACAATCTACTAAATTCAAGATAATATATTTTAGGTATAATGGTT

[0961] ACTTCACAAATAATATTAATTAATTAATTACTTATAAACTAAAATTTAATTCAAAACTATG

[0962] TTTAATTTAACCAATTCTCTCGTGCCTAAATTATATCTTTTTAAACTTTTGTATTAAGTT

[0963] GCATTTTAAACTTATAAAACACATTCAATACACATATTATCGATGTGTATCAAACTAATA

[0964] GTCGTAAAGGTGCATTGTTGCCAACATGAGTACAGCTGAACTGACATAATATTTGTATA

[0965] TATGAAGTTCTATTAATGTGCCTAAATATTGATTAATGGTTATTATACAATTTCTTTTAC

[0966] CTATATTCTATATTTTTCTATATTTACGATTGTTGTTTTAGCCACTATTTTTATCTAATT

[0967] TGTTTTGTATGGCAATTACATACCCCCTTTTCCTTTTGGTTTTTTTTTTTTTCAATGTG

[0968] TTTTAGGGTTGTGAAAGGTTGAAGCATGTTGATCTTAGCTACTCTCCGTTATTAGAGAAA

[0969] ATTCCTAATTTGTCTGCAGCATCAAACCTTGAACAGTTGTATCTCATAAATTGCAAAAAT

[0970] TTAGGAATGATAGATAAGTCTGTTTTTTCTCTCGATAAGCTTAATGTCCTAAACCTTGAT

[0971] GGTTGTTGTAACCTTCAAAAGCTTCCAAGAGGCTATTTCATGTTAAGTTCTCTCGGTAT

[0972] TTGAATCTCTCTTACTGCAAAAAGCTTGAAAAAAATTCCAGACTTATCTGCAGCATCAAAC

[0973] CTTAAAAGATTGTATCTCCAAGAATGCACAAATTTAAGAGTGATTCATGAATCTGTTGGA

[0974] TCTTTGGATAAGCTTGATCATCTGGACCTTAGACAATGCACTAAACTGGTAAAGCTTCCA

[0975] AGCTATCTCAGGTTAAAGTCTCTTGAATATTTATCACTTTCTGGGTGTTGTAAGCTTGAA

[0976] AGCTTCCCCAACAATTGCTGAAAACATGAAATCTTTATGGGGATTGGATTTGGATTTTACT

[0977] GCCATAAAGGAGTTACCTTCATCAATTGGATATCTTACTAAGCTTTCTATATTAAAACTT

[0978] AACGGTTGCACAAACCTCATCTCCCTTCCCAATACAATTTATTTGTTAAGGAATCTTGAG

[0979] AATCTTCTTCTTAGTGGCTGTTCTATATTTGGAATGTTTCCCCATACATGGGACCCAACC

[0980] ATCCCAACCATCCAACAAGTATGCTCTCCTTCAAATTGATGGAAACAGCTTCCTGGAGC

[0981] TTAGAATTTCCGATTTACTAGTACCAAATGAAAGTTTATGTGCACATTTCACTTTGTTG

[0982] GATCTTGAATCTTGCAACATATCAAATGCAAAATTTTTAGAATTATTGTGTGATGTTGCC

[0983] CCTTTCTTATCTGATCTACGCTTGTCTGAAAACAAATTCTCTAGTTTACCCTCATGTCTC

[0984] CACAAGTTCATGTCCTTGTGGAATCTTGAATTAAGGAATTGCAAGTTTCTTCAAGAAATT

[0985] CCAAACCTTCCTGAGAATATACAAAAAATGGATGCCAGTGGTTGTGAATCGTTGGCTCGA

[0986] AATCCAGATAACATTGTGGATATAATATCAAAAAAACAGGTTCGCCTCTAATTTCCATTC

[0987] AATTTATATTCTTATCTTGTAAACAATTTAATGCATTATGAATTCTTGTTCTCTATAGGA

[0988] CCTCACATTGGGTGAGATTTCAAGAGAGGTTTTTATTAACGGGGATTGAGATTCCAGAATG

[0989] GTTCAGCTATAAGACTACATCCAATTTAGTGACAGCTAGCTTTCGTCACTATCCAGACAT

[0990] GGAAAGAACTTTGGCTGCCTGTGTTAGTTTTCAAAGTGAATGGAGATTCATCTAAAAGAGG

[0991] AGCCCAAATGTCATGTAGTATATTCATCTGCAGTAAACTCCATTCTTCATTTTCAAGACC

[0992] ATTTCTTCCATCAAAATCAGAATATATGTGGTTAGTAACAACTTCTCTAGCGTGGGGGTTC

[0993] CATGGAGGTGAATGATTGGAATAAAGTTTTGGTCTGGTTTGAGGTTCATGAAGCACATAG

[0994] TGAGGTTAATGCAACTATAACAAGGTGTGGTGTTCATGTCACTGAAGAGCTCCATGGGAT

[0995] ACAAATGGATGTCAAGTGGCCGATGGTAAATTATGCTGATTTTTATCAACTGGAGAAATT

[0996] GCAAAGTCTGTAAGTTGATTGTTTACTTGTTATTTATTTAGTCTCTTTCTTTTTCTTTTT

[0997] TTTTTTTTTTTTTTTTTTTTTGGAGTGAAAGTATATAATAGATCTCAAAGGGGGAGATGAGT

[0998] ATGTTATGATTTTGTTTGTTTAATGGCATAGGGATATTGAGGATCTTCTTCTCAAAAGCT

[0999] TTTTAGAAACAGTCTCTTGCCTGTCAAATTCCAAAGCAGCAATGTTACATGCAGGAAATT

[1000] ATGATCCAGAAGCAATAATTGATTCCAACATACAACCTATGATTTTCCCATTGCACGTAA

[1001] CATATAATGGTGAAACAGTGATATGTGGAATGGAAGGCATGGGAGATACTACACTCGCCA

[1002] ACTCTTTATGCAATAAATTTAAAGGGATGGAGGGCCAATGCGGTGAAGCTTTAGATAATT

[1003] CTACAAGCTTTTTCCATATCAAAAGAAGACAACTCCTGAGCTATTCCTGGTCGCCGGCGG

[1004] TCCACCATCGTAAGTGTGGAGATGGTGAAAGAGGAACGAATATCACAACCCACACAATAT

[1005] CCTCCAAACGCTATTTGATACTCCTTCGTGAAGCCAAGAGCTATCAGGATGTACATGACT

[1006] GGTTTTATACACATTGTTGGATAAAAGCTTCATATTGCAGTTATGACGGAAGAGGTGATG

[1007] GTGTGTTTCTGATTAAAGGGGTTGATACATCCTTGCTCTGA

[1008] SEQ ID NO 14: cDNA (mRNA) encoding SEQ ID No: 9

[1009] ATGGGTTCTTCTGCTCTTCCAGTTGAATCTTCTTCT

[1010] TCTTCTTCTTCTTCTTCTCCCAACTTTCTCTATTACTACGATTATGATGTGTTTTTTTAGT

[1011] TTCAGGAGGAGAAGACACTCGCTCCAGTTCATCAGTCATCTTCATATGGCCTTGCGTCTA

[1012] AAGGGAGTCAACGTCTTCATAGACGACAAACTCAAAAGGGGTGACCAAATCTCTGAGTCT

[1013] CTTTCTAAATCTATAGAGCGATCTAGACTTTCACTCGTTATTTTCTCTAAAAATTATGCA

[1014] TCTTCAACTTGGTGTTTGGATGAACTGGTGAAAATAATTGAGTGTATGAAACCCAAAGGA

[1015] CAGCTAGTTTTGTCTGTCTTCTACAAGGTGGATCCGTCCGTGGTTTGGAAACAAACAGGT

[1016] GGGTTTGGTGAAGCATTGGCCAAACATGAGGCTACTAAGTTAACAACCAACAAGATTAAA

[1017] CCATGGAAGGAAGCTTTGACTATTGCTGCTGGTTTGTCTGGTTGGGATCTAGCAAATTGC

[1018] AAGGATGAGGCTGAACTTATCCAAGAAATTGTTAAACGAGTGTTGTCTGTATTAAATCCA

[1019] ATGCAATTACTACATGTAGCCAAGCATCCAGTTGGAATTGATTTTCGATTAAGGAAAATT

[1020] GAGGAGTTGGTCTCTCATATTGGGTCCGAGGGTGTTAACATGGTGGGGATGTATGGCATT

[1021] GGAGGCATTGGTAAGACCACTTTGGCTAAGGCTTTGTACAACAAAATTGCTAACCAATTT

[1022] GAAGGATCTTGCTTTCTACAAGATGTTAGACGAGAAGCTTCAAAGCATGGGCTCGTTAAA

[1023] CTACAAGAAACCTTACTCCATGACATCTTAAAGGAGGATTTGAAGGTTGTCAGCCGTGAT

[1024] AGAGGAATTAACATCATAAGGAGTAGACTGTGTTCAAAGAAAGTTCTTATAGTTCTTGAT

[1025] GATGTGGATGATCGTGAGCAATTAGAAGCACTGGTTGGTGGTCGTGATTGGTTTGGTCGA

[1026] GGTAGCAAAATCATTGTGACGACAAGGAATGAGCATTTACTTTTTAGCCATGGATTTGAT

[1027] GATCAAAAGCATAAAATTCAAGAATTGAATCAAGATCATGCTCTTGAGCTTTTTAGTTGG

[1028] CACGCTTTTAAGAAAAGCCATCCATCAAGTAATTATCTAGGCCTTTCAGAACGTGCTACA

[1029] AATTATTGTAAAGGTCTATCTTTGGCACTCGTTGTTTTGGGTTCTTTCCTTCGTGGCAGA

[1030] GATCAAGCAGAATGGAACTGTATATTAGATGAATTTGAAACCTCTTTGAGAAAAGATATT

[1031] AAAGATGTTCTTCAATTAAGTTTTGATGGACTTGAAGACAAAGCAAAGGAAATTTTCCTT

[1032] GATATTTCTTGTTTACTCGTGGGAGAAGAAATCAATTGTGCTAAAAAGATGTTGAGTGCA

[1033] TGCCATTTGAACATAGATTTTGAAATTATGATACTCATGGATTCTTTCACTTGTTACTATT

[1034] GAAACGGATAGAGTGCAAATGCATGAGTTAATACAACAAATGGGTCGTAGCATAGTTCAT

[1035] AATGAATCATCTGAGCCTGGAAAGAGGAGTAGGTTGTGGTTGGTGCAGGACATTTGGGAG

[1036] GTGTTTGTTAATAATTCAGGAACAGATGCAGTTAAAGCCATAAAGTTGGACTTGCCTAAT

[1037] CCCACAAAGCTAAATGTAGATCCACAAGCATTTAGAAGTATGAAAAATTTGAGATTGCTT

[1038] ATCATTCGAAATGCACAATTTTGTAGAAAGATTAAGTACCTACCTAATAGCTTAAAGTGG

[1039] ATTGAGTGGCGTGGATTTGCTCATCGATCTTTGCCGTCATGCTTCATTACCAAAATCTT

[1040] GTTGGACTTGATTTGCGACATAGCTCCATCAAGATTTGGGAAAAGGCTCAAGGGTTGT

[1041] GAAAGGTTGAAGCATGTTGATCTTAGCTACTCTCCGTTATTAGAGAAAATTCCTAATTTG

[1042] TCTGCAGCATCAAACCTTGAACAGTTGTATCTCATAAATTGCAAAAATTTAGGAATGATA

[1043] GATAAGTCTGTTTTTTCTCTCGATAAGCTTAATGTCCTAAACCTTGATGGTTGTTGTAAC

[1044] CTTCAAAAGCTTCCAAGAGGCTATTTCATGTTAAGTTCTCTTCGGTATTTGAATCTCTCT

[1045] TACTGCAAAAAGCTTGAAAAAATTCCAGACTTATCTGCAGCATCAAACCTTAAAAGATTG

[1046] TATCTCCAAGAATGCACAAATTTAAGAGTGATTCATGAATCTGTTGGATCTTTGGATAAG

[1047] CTTGATCATCTGGACCTTAGACAATGCACTAAACTGGTAAAGCTTCCAAGCTATCTCAGG

[1048] TTAAAGTCTCTTGAATATTTATCACTTTCTGGGTGTTGTAAGCTTGAAAGCTTCCCAACA

[1049] ATTGCTGAAAACATGAAATCTTTATGGGGATTGGATTTGGATTTTACTGCCATAAAGGAG

[1050] TTACCTTCATCAATTGGATATCTTACTAAGCTTTCTATATTAAAACTTAACGGTTGCACA

[1051] AACCTCATCTCCCTTCCCAATACAATTTATTTGTTAAGGAATCTTGAGAATCTTCTTCTT

[1052] AGTGGCTGTTCTATATTTGGAATGTTTCCCCATACATGGGACCCAACCATCCCAACCATC

[1053] CAACAAGTATGCTCTCCTTCAAAAATGATGGAAACAGCTTCCTGGAGCTTAGAATTTCCC

[1054] GATTTACTAGTACCAAATGAAAGTTTATGTGCACATTTCACTTTGTTGGATCTTGAATCT

[1055] TGCAACATATCAAATGCAAAATTTTTAGAATTATTGTGTGATGTTGCCCCTTTCTTATCT

[1056] GATCTACGCTTGTCTGAAAACAAATTCTCTAGTTTACCCTCATGTCTCCACAAGTTCATG

[1057] TCCTTGTGGAATCTTGAATTAAGGAATTGCAAGTTTCTTCAAGAAATTCCAAACCTTCCT

[1058] GAGAATATACAAAAAATGGATGCCAGTGGTTGTGAATCGTTGGCTCGAAATCCAGATAAC

[1059] ATTGTGGATATAATATCAAAAAAACAGGACCTCACATTGGGTGAGATTTCAAGAGAGTTTT

[1060] TTATTAACGGGGATTGAGATTCCAGAATGGTTCAAGCTATAAGACTACATCCAATTTAGTG

[1061] ACAGCTAGCTTTCGTCACTATCCAGACATGGAAAGAACTTTGGCTGCCTGTGTTAGTTTC

[1062] AAAGTGAATGGAGATTCATCTAAAAGAGGAGCCCAAATGTCATGTAGTATATTCATCTGC

[1063] AGTAAACTCCATTCTTCATTTTCAAGACCATTTCTTCCATCAAAATCAGAATATATGTGG

[1064] TTAGTAACAACTTCTCTAGCGTGGGGTTCCATGGAGGTGAATGATTGGAATAAAGTTTTG

[1065] GTCTGGTTTGAGGTTCATGAAGCACATAGTGAGGTTAATGCAACTATAACAAGGTGTGGT

[1066] GTTCATGTCACTGAAGAGCTCCATGGGATACAAATGGATGTCAAGTGGCCGATGGTAAAT

[1067] TATGCTGATTTTTATCAACTGGAGAAATTGCAAAGTCTGGATATTGAGGATCTTCTTCTC

[1068] AAAAGCTTTTTAGAAACAGTCTCTTGCCTGTCAAATTCCAAAGCAGCAATGTTACATGCA

[1069] GGAAATTATGATCCAGAAGCAATAATTGATTCCAACATACAACCTATGATTTTCCCATTG

[1070] CACGTAACATATAATGGTGAAACAGTGATATGTGGAATGGAAGGCATGGGAGATACTACA

[1071] CTCGCCAACTCTTTATGCAATAAATTTAAAGGGATGGAGGGCCAATGCGGTGAAGCTTTA

[1072] GATAATTCTACAAGCTTTTTCCATATCAAAAGAAGACAACTCCTGAGCTATTCCTGGTCG

[1073] CCGGCGGTCCACCATCGTAAGTGTGGAGATGGTGAAAGAGGAACGAATATCACAACCCAC

[1074] ACAATATCCTCCAAACGCTATTTGATACTCCTTCGTGAAGCCAAGAGCTATCAGGATGTA

[1075] CATGACTGGTTTTATACACATTGTTGGATAAAAGCTTCATATTGCAGTTATGACGGAAGA

[1076] GGTGATGGTGTGTTTCTGATTAAAGGGGTTGATACATCCTTGCTC

[1077] SEQ ID NO 15: FAM primers for KASP determination of SNP_01

[1078] GAAGGTGACCAAGTTCATGCTCGGTAGTATTTCTTTTGGTAATATTTGTAC

[1079] SEQ ID NO 16: VIC primers for KASP determination of SNP_01

[1080] GAAGGTCGGAGTCAACGGATTCGGTAGTATTTCTTTTGGTAATATTTGTAT

[1081] SEQ ID NO 17: Common primers for KASP determination of SNP_01

[1082] CGGCTAATAAAGCAGCTTCATCATCTAAA

[1083] SEQ ID NO 18: FAM primers for KASP determination of SNP_02

[1084] GAAGGTGACCAAGTTCATGCTAAATTACACCTTCTGGAACCTTTGAAATT

[1085] SEQ ID NO 19: VIC primers for KASP determination of SNP_02

[1086] GAAGGTCGGAGTCAACGGATTACACCTTCTGGAACCTTTGAAATC

[1087] SEQ ID NO 20: Common primers for KASP determination of SNP_02

[1088] CTTTATACAATGGATTGTGAGTTGCTYGAA

[1089] SEQ ID NO 21: FAM primers for KASP determination of SNP_03

[1090] GAAGGTGACCAAGTTCATGCTGTCAAAGTTAATTCTTTAGTTAATGAGATAAA

[1091] SEQ ID NO 22: VIC primers for KASP determination of SNP_03

[1092] GAAGGTCGGAGTCAACGGATTGTCAAAGTTAATTCTTTAGTTAATGAGATAAG

[1093] SEQ ID NO 23: Common primers for KASP determination of SNP_03

[1094] GATGTGGGTCCCTGTTTTCGTTCTT

[1095] SEQ ID NO 24: FAM primers for KASP determination of SNP_04

[1096] GAAGGTGACCAAGTTCATGCTCTCAAGATGCCATTTAATTGTGTGTA

[1097] SEQ ID NO 25: VIC primers for KASP determination of SNP_04

[1098] GAAGGTCGGAGTCAACGGATTCTCAAGATGCCATTTAATTGTGTGTG

[1099] SEQ ID NO 26: Common primers for KASP determination of SNP_04

[1100] CCCTCCTGTTGTCAAGTACATAATCATAT

[1101] SEQ ID NO 27: FAM primers for KASP determination of SNP_05

[1102] GAAGGTGACCAAGTTCATGCTTGTAGAAATTGTCAAAACTAAATACAGTAAT

[1103] SEQ ID NO 28: VIC primers for KASP determination of SNP_05

[1104] GAAGGTCGGAGTCAACGGATTGTAGAAATTGTCAAAACTAAATACAGTAAC

[1105] SEQ ID NO 29: Common primers for KASP determination of SNP_05

[1106] CGTGAGATGGGCCATATTGTAATATAACAT

[1107] SEQ ID NO 30: FAM primers for KASP determination of SNP_06

[1108] GAAGGTGACCAAGTTCATGCTCCCTTTATTTCTCTCCGTCAGTC

[1109] SEQ ID NO 31: VIC primers for KASP determination of SNP_06

[1110] GAAGGTCGGAGTCAACGGATTCCCCTTTATTTCTCTCCGTCAGTT

[1111] SEQ ID NO 32: Common primers for KASP determination of SNP_06

[1112] CTTATCAAATAAAATTGGGGAGTAGGAGAT

[1113] SEQ ID NO 33: FAM primers for KASP determination of SNP_07

[1114] GAAGGTGACCAAGTTCATGCTAGAACTCTTCTACATTCCCCTCC

[1115] SEQ ID NO 34: VIC primers for KASP determination of SNP_07

[1116] GAAGGTCGGAGTCAACGGATTGAGAACTCTTCTACATTCCCCTCT

[1117] SEQ ID NO 35: Common primers for KASP determination of SNP_07

[1118] GGAGAAGGAAGTAGGTGTGTCTGAT

[1119] SEQ ID NO 36: Genomic DNA encoding the susceptibility protein of SEQ ID NO: 10

[1120] ATGGGTTCTTCTGCTCTTCCAGTTGAATCTTCTTCT

[1121] TCTTCTTCTTCTCCCAACTTTCTCTATTACTACGATTATGATGTGTTTTTTAGTTTCAGA

[1122] GGAGAAGACACTCGCTCCAGTTTCATCAGTCATCTTCATATGGCCTTGCGTCTAAAGGGA

[1123] GTCAACGTCTTCATAGACGACAAACTCAAAAGGGGTGACCAAATCTCTGAGTCTCTTCTC

[1124] AAATCTATAGAGCGATCTAGACTTTCACTCGTTATTTTCTCTAAAAATTATGCATCTTCA

[1125] ACTTGGTGTTTGGATGAACTGGTGAAAATAATTGAGTATAAAAAATCCAAAAGTCAAGCG

[1126] GTTTTGCCGGTGTTCTACAAGGTGGATCCGTCCGAGGTTCGAAAAAACCGGTGGGTTT

[1127] GGGGAAGCATTAGCCAAACATGAAGCTAATAAGTTATTGACCAACAAGATTCAACCATGG

[1128] AAGGAAGCTTTGACTTTTGCTGCTGGTTTGTCTGGTTGGGATCTAGCAAATTGGTATTTC

[1129] TTTTTTTAATTTCCCAAGACTCATTGTCTAAGTGAAGTTTAAATTTATCTACTATTTCCT

[1130] ATTTTCATTTTTTTTTTGTATGATGTTACTTTACAACAGCAAGGATGAGGCTGAACTTA

[1131] TCCAAGAAATTGTTAAACGAGTGTTGTCTGTATTAAATCCAATGCAATTACTACATGTAG

[1132] CCAAGCATCCAGTTGGAATTGATTTTCGATTAAGGAAAATTGAGGAGTTGGTCTCTCATA

[1133] TTGGGTCCGAGGGTGTTAACATGGTGGGGATGTATGGCATTGGAGGCATTGGTAAGACCA

[1134] CTTTAGCTAAGGCTTTGTACAACAAAATTGCTAACCAATTTGAAGGATGTTGCTTTCTAC

[1135] AAGATGTTAGACGAGAAGCTTCAAAGCATGGGCTCGTTAAACTACAAGAAACCTTACTCA

[1136] ATGACATCTTAAAGGAGGATTTGAAGGTTGTCAGCCGTGATAGAGGAATTAACATCATAA

[1137] GGAGTAGACTGTGTTCAAAGAAAGTTCTTATAGTTCTTGATGATGTGGATGATCGTGAGC

[1138] AATTAGAAGCACTGGTTGGTGGTCGTGATTGGTTTGGTCGAGGGTAGCAAAATCATTGTGA

[1139] CGACAAGGAATGAGCATTTACTTTTTAGCCATGGATTTGATGATCAAAAGCATAAAATTC

[1140] AAGAATTGAATCAAGATCATGCTCTTGAACTTTTTAGTTGGCACGCTTTTAAGAAAAGCC

[1141] ATCCATCAAGTAATTATCTAGGCCTTTCAGAACGTGCTACAAATTATTGTAAAGGTCTAT

[1142] CTTTGGCACTCGTTGTTTTGGGTTCTTTCCTTCGTGGCAGAGATCAAGCAGAATGGAACT

[1143] GTATATTAGATGAATTTGAAACCTCTTTGAGAAAAGATATTAAAGATGTTCTTCAATTAA

[1144] GTTTTGATGGACTTGAAGACAAAGCAAAGGATATTTTCCTTGATATTTCTTGTTTACTTG

[1145] TGGGAGAAGAATACAATTGTGCTAAAAAAATGTTGAGTGCATGCCATTTGAACATAGATT

[1146] TTGGAATTATGATACTCGTGGATCTTTCACTTGTTACTATTGAAACGGATAGAGTGCAAA

[1147] TGCATGAGTTAATACAACAAATGGGTCGTAGCATAGTTCATAATGAATCATCTGAGCCTG

[1148] GAAAGAGGAGTAGGTTGTGGTTGGTGCAGGACATTTGGGAGGTGTTTGTTAATAATTCAG

[1149] TGAGTAACTCCTACCTAAAGTATTTAATAATTTGGACTTCACCCGATAATATATAAATGT

[1150] TTGTCAAAGTAATTAAATTGTCAGTAATGTTAAATTACTAAATATATGACTTTGTAGGGA

[1151] ACAGATGCAGTTAAAGCCATAAAGTTGGACTTGCCTAATCCCACAAAGCTAAATGTAGAT

[1152] CCACAAGCATTTAGAAGCATGAAAAATTTGAGATTGCTTATCATTCGAAATGCACAATTT

[1153] TGTAGAAAGATTAAGTACCTACCTAATAGCTTAAAGTGGATTGAGTGGCGTGGATTTGCT

[1154] CATCGATCTTTGCCGTCATGCTTCATTACCAAATCTTGTTGGACTTGATTTGCGACAT

[1155] AGCTCCATCAAAAGATTTGGGAAAAGGCTCGAGGTAAAAATATATTTCTTTTTGCATCTGT

[1156] ATTTGACTGGAGATTTCTTCATTGATTTTCTTAAAAAGTTTTAATGGGTAGCTTAACTGC

[1157] ACCGGAAAAAAAAAATCACAAATAGGTGATTTAGCAAAATTTAACATGGATTAACCCGTG

[1158] CTAATCACATACTTTTAAAGAATTTGCAAATTATATCAAAGTTTATTAATGATATAGTGA

[1159] TCTATCGCACAACCATAGACTTATGCTAGTCATATGATCTATCATTAATATACTCCTATT

[1160] AGTGATATGATCTACATGGATTAATTCCAAGGGGTCTTTTGGGGAAAGGGTTGGATTATG

[1161] GGAGTTAAGGTTATGATAAAACTAGTGTTATGACAAATCTAGGATTACGATAAAGATGTA

[1162] TTTAGTGGAAGGGTTATGGAGGAAGTATTATGATAAAATATGTTTGGGAGAAGGGTTATG

[1163] TGGGTAGGGTTATGATAAATATATATATATATATATATATATATATATATATATATATAT

[1164] ATATTAATTCATATTATAAATCAAATACACAAATTTCGTATATTTAATTTACTAAATTGT

[1165] CCTAGTTTTCATAAAATAATTTGCCTTAATTTATTTAACTACGTTATTCAATTTCAACAT

[1166] ACACATTGTTTATTTTCAACATTTTTTAAGTATACGTTACGTTTCAATAATTTGGTTTTC

[1167] ATGAATATGACACACAACCTGTAAATATGTAAATAAGGGTAAATAACAATTGAAAATGCG

[1168] AATTACCGAAAACAGAAAATAATAATCAACGCTTTTACAAACTCTAGTTTTGTTTTCTAG

[1169] TGAAAAATTACGATGAATCTAGTTTTAGGTTTCTCTTCAATTTATTTTAGTCTACTTTTA

[1170] AAATATTAACTTTATTTTTTATTCTTTTTAACGGTGATTTATACTTTCAAAATATTTATT

[1171] TTAGTCATTTTGCAACAATTATTGGGTAAAAAATTCATGAATCTACCGCCTCTATAACAA

[1172] AAAAATTTGTTATATTAATTTAACCAAGCTTAATTTAATAACCAAATTAACATTATGAAA

[1173] GTGGGAAAATATAGACAAACTGCATCTAAATAATAGATTTGTTAAATTATAGCCCCACCA

[1174] ATTTTTAATTATTCAAATAGTCCTTATGATTTTCTAATTGTTTTAATCAATTTATGGTA

[1175] GTTTGTTAAAATTTTACCATTTTTTTTATTTAATTGATATTCCTTAGATATTTATCCTAG

[1176] CTAACTAAAAAAACAATGAGATTGAGATTTATATTAAAATTTTAATTTCATAAATATA

[1177] TTGATATGAATTCAAACATAGAGTTGATTGAATGTAACTAAATTAAAAATTTAATTGTGA

[1178] AATCACTATAAATTATCTAATTCAAGAGTAAAGGAAAAAATTTAAATTTTAATAATGTAA

[1179] TTATACCAAATTCATAATTAAAAAACAAGCTGTGTTTTTAAAATAAAAAGATTAAAATGA

[1180] ACAAAACTCATAAATATAAAAATTATAAATGAGAAAAAAATGAATTGAGGAAGGGTTGAA

[1181] GAAAGGTTGAGGGGATAAAACTAAGGTTATGATAACCCTATAATTAAAGACTAAACTA

[1182] CATTCTCATTCTGTTTAAGGTATCAAATTAAATGGTTAGTTTACAATCTACTAAATTCAA

[1183] GATAATATATTTTAGGTATAATGGTTACTTTACAAATAATATTAATTAATTAATTACTTAT

[1184] AACTAAAATTTAATTCAAAACTATGTTTAATTTAACTAATTCTCTCGTGCCTAAATTAT

[1185] ATTTTTTAAACTTTTGTATTAAGTTGCATTTTAAACTTATAAACACACTTACTATCGA

[1186] TATGTATCAAACTAATAGTTGTAAAAAGTGCATTGTTGCTAACATGAGTACAGCTGAACT

[1187] GACATAATATTGATTAATGGTTATTATACAATTTCTTTTACCTATATTCTATATTTTTCT

[1188] ATATTTACGATTGTTGTTTTAGCCACTATTTTATCTAAATTTTGTTTTGTATGGCAATTA

[1189] CATACCTCCTTTTCCTTTTGGTTTTTTTTTTTTCAATGTGTTTTAGGGTTGTGAAAGGTT

[1190] GAAGCATGTTGATCTTAGCTACTCTACTTTATTAGAGCAAATTAATGATTTCTCTCCGGC

[1191] ATCAAATCTTGAAGAATTGCATCTCATCAATTGCACAAATTTAGGAATGATAGATAAGTC

[1192] TGTTTTTTCTCTCTAAGCTTAGTTCCTAAACCTTGATGGTTGTTGTAACCTTCAAAA

[1193] GCTTCCAAGAGGCTATTTCATGTTAAGTTCTCTTAAAGAATTGAATCTCTGTTACTGCAA

[1194] AAAGCTTGAAAAAATTCCAGACTTATCTGCAGCATCAAACCTTAAGAGATTGTATCTCCA

[1195] AGAATGCACAAATTTAAGAGTGATTCATGAATCTGTTGGATCTTTGGATAAGCTTAATCA

[1196] TTCTGGACCTTAGACAATGCACTAAACTGGTAAAGCTTCCAAGCTATCTCAGGTTAAAGTC

[1197] TCTTTCCAATTTATTACTTTCTGGGTGTTGTAAGCTTGAAAGCTTCCCAACAATTGCTGA

[1198] AAACATGAAATCTTTAAGGGAATTGGATATGGATTTTACTGCCATAAAGGAGTTACCTTC

[1199] ATCAATTGGATATCTTACTAACCTTTCTATATTAAAACTTAACGGTTGCACAAACCTCAT

[1200] CTCCCTTCCCAATACAATTTTATTGTTAAGGAATCTTTGAGAATCTTCTTCTTAGTGGCTG

[1201] TTCTATATTTGGAATGTTTCCCCATACATGGGACCCAACCATCCCAACCATCCAACAAGT

[1202] ATGCTCTCCTTCAAAAATGATGGAACAGCTTCCTGGAGCTTAGAATTTCCCCATTTACT

[1203] AGTACCAAATGAAGTTTATGTGCCCATTTCACTTTGTTGGATCTTGAATCTTGCAACAT

[1204] ATCAAATGCAAAATTTTTAGAATTATTATGTGATGTTGCCCCTTTCTTATCTGATCTACG

[1205] CTTGTCTGAAAACAAATTCTCTAGTTTACCCTCATGTCTCCACAAGTTCATGTCCTTGTG

[1206] GAATCTTTGAATTAAGGAATTGCAAGTTTCTTCAAGAAATTCCAAACCTTCCTGAGAATAT

[1207] ACAAAAAATGGATGCCAGTGGTTGTGAATCGTTGGCTCGAAATCCAGATAACATTGTGGA

[1208] TATAATATCAAAAAAACAGGTTCGCCTCTAATTTCCATTCAATTTATATTCTTATCTTGT

[1209] AAACAATTTAATGCATTATGAATTCTTGTTCTCTATAGGACCTCACATTGGGTGAGATTT

[1210] CAAGAGAGTTTTTTATTAACGGGGGATTGAGATTCCAGAATGGTTCAGCTATAAGACTACAT

[1211] CCAATTTAGTGACAGCTAGCTTTCGTCACTATCCAGACATGGAAAGAACTTTGGCTGCCT

[1212] GTGTTAGTTTCAAAGTGAATGGAGATTCATCTAAAAAGAGGAGCCCAAATTTCATGTAGTA

[1213] TATTCATCTGCAGTAAACTCCATTCTTCATTTTCAAGACCATTTCTTCCATCAAAATCAG

[1214] AATATATGTGGTTAGTAACAACTTCTCTAGCGTGGGGGTTCCATGGAGGTGAATGATTGGA

[1215] ATAAAGTTTTGGTCTGGTTTGAGGTTCATGAAGCACATAGTGAGGTTAATGCAACTATAA

[1216] CAAGGTGTGGTGTTCATGTCACTGAAGAGCTCCATGGGATACAAATGGATGTCAAGTGGC

[1217] CGATGGTAATTATGCTGATTTTTATCAACTGGAGAAAATTGCAAAGTCTGTAAGTTGATT

[1218] GTTAACTTGTTATTTATTTATTCTCTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTG

[1219] GAGTGAGAGTATATAATAGATCTCAAAGGGGGAGATGAGTATGTTATGATTTTGTTTGTT

[1220] TAATGGCATAGGGATATTGAGGATCTTCTCTCAAAAGCTTTTTAGAAACAGTCTCTTGC

[1221] CTGTCAAATTCCAAAGCAGCAATGTTACATGCAGGAAATTATGATCCAGAAGCAATAATT

[1222] GATTCCAACATACAACCTATGATATTTCCATTGCACGTAACAAATAATGATGGCACATAT

[1223] ATATGTGGAGGCATGGGAGGCACTGCACTTGCCAACTCTTTATGCAATAAATTTAAAGGG

[1224] ATGGAGGGCCAATGCGGTGAAGCTTTAGATAATTCTACAAGCTTTTTCCATATCAAAAGA

[1225] AGACAACTCCTGAGCTATTCCTGGTCGCCGGCGGTCCACCATCGTAAGTGTGGAGATGGT

[1226] GAAAGAGGAACGAATATCACAACCCACACAATATCCTCCAAACGCTATTTGATACTCCTT

[1227] CGTGAAGCCAAGAGCTATCAGGATGTACATGACTGGTTTTATACACATTGTTGGATAAAA

[1228] GCTTCATATTGCAGTTATGACGGAAGAGGTGATGGTGTGATTCTGATTGAAGGGGTTGAT

[1229] ACATCCTTGCTCTGA

[1230] General methods

[1231] 1. Determine the symptom level of plants infected with ToLCNDV-ES.

[1232] 1.1 Plants and pathogens (viruses)

[1233] Melon plants were infected with the ToLCNDV-ES strain that infects melon plants. In this invention, the invasive ToLCNDV-ES isolate named MU_13_002, isolated in Almería, Spain, was used as the inoculum.

[1234] 1.2 ToLCNDV-ES propagation

[1235] The ToLCNDV-ES inoculum is kept on live, infected melon plants. It is essential to ensure that pure virus isolates are used and that neither the virus source nor the whiteflies are contaminated with other diseases, especially other viruses such as CGMMV, CYSDV, and CYVY SqMV. To pre-propagate the ToLCNDV-ES inoculum, whiteflies (tobacco whiteflies) are fed to ToLCNDV-ES-sensitive (susceptible) infected melon plants in insect validation cages. Before infecting test plants, ToLCNDV-ES-infected plants are placed in the insect validation cages, and whiteflies are released into the same cages and fed to ToLCNDV-ES-infected plants for approximately 3 days.

[1236] 1.3 Inoculation of the plants to be tested

[1237] For each genotype of melon plants to be analyzed, 18 plants were grown until the first true leaf unfolded (usually 12-15 days after sowing), with 15 plants infected and 3 plants simulantly infected. Also included were 15 plants of susceptible varieties; in this experiment, susceptible varieties were Vedantrais and susceptible Galia and Piel de Sapo. The 15 plants of each genotype for testing ToLCNDV-ES resistance were placed in insect cages, and infected whiteflies obtained as described in 1.2 above were released into the cages to infect the plants. It was essential to ensure that each test plant in the cage had at least 5-10 whiteflies available. The whiteflies and test plants were kept in the cages for approximately 48 hours, and then the whiteflies were eliminated with an appropriate insecticide. Three plants of each genotype were also used for simulant infection, treated in the same manner as the test plants except that the whiteflies used for infection did not contain ToLCNDV-ES. Three replicate settings were created, with 5 plants inoculated and 1 simulant inoculated for each genotype.

[1238] 1.4 Inducing the growth of infected test plants

[1239] The infected test plants obtained as described in 1.3 were transplanted into larger pots and placed in a greenhouse equipped with cooling facilities. The plants were grown at nighttime temperatures of approximately 18°C ​​and daytime temperatures of approximately 25°C, with daylight hours ranging from 14 to 16 hours. Infected plants of each infection genotype were grown in triplicate, with each replicate containing five ToLCNDV-infected plants and one simulated infected plant. These plots were randomized relative to the growth area.

[1240] 1.5. Scoring the symptom levels of ToLCNDV-ES infection.

[1241] Symptom level scoring may have been performed approximately 25 days (dpi) after ToLCNDV-ES infection, but is preferably performed approximately 30 or 40 days (dpi) or later. In the presence of plants showing recovery from viral infection, further symptom scoring may be performed approximately 45 or 55 days (dpi) after ToLCNDV-ES infection.

[1242] Based on the phenotypes shown below, use the following symptom levels:

[1243] Symptom level Observed phenotypes 1 Plant death 2 Severe mosaic virus and leaf curling, yellowing, and stunted growth. No recovery. 3 Severe mosaic virus and leaf curling, yellowing, and stunted growth. No recovery. 4 Leaf curling and mosaic virus, chlorosis, no growth decline or slight growth reduction. No recovery. 5 Leaf curling and mosaic virus, chlorosis, and no growth decline. The upper part of the plant shows slight recovery. 6 Mild leaf curling, mosaic virus, and chlorosis; no growth decline. The upper and middle regions of the plant have recovered. 7 Mild leaf curling, mosaic virus, and chlorosis; no stunted growth. Symptoms appear only in the lower part of the plant. 8 Mild mosaic virus 9 Asymptomatic

[1244] 1.6 Optional Additional Tests

[1245] It is recommended to use at least one genotype highly resistant to ToLCNDV-ES (symptom level 8-9) and one genotype highly susceptible to ToLCNDV-ES (symptom level 1) in each experimental setup. It is further recommended to also include a genotype with moderate resistance to ToLCNDV-ES infection in each test setup. Optimal results are obtained when the aforementioned genotypes are included, and the symptom level of each genotype is scored relative to the results obtained with highly resistant, highly susceptible, and moderately resistant genotypes. These genotypes also provide a clear indication of the ToLCNDV-ES infection level of whiteflies in melon plants.

[1246] In addition, it is recommended to examine the infection and spread of ToLCNDV-ES in both infected and control plants. This can be done by examining the presence and amount of viral DNA in the upper parts of the plant. A suitable way to examine the presence and amount of ToLCNDV-ES DNA in the upper plant parts is to hybridize the plant material with a probe that hybridizes with the DNA of the ToLCNDV-ES strain used. Various transformation techniques are well known in the art. Simple so-called dot blot analysis is sufficient to obtain valuable results. Similarly, PCR or quantitative PCR techniques can be used.

[1247] Example

[1248] Example 1 - Selection of ToLCNDV-ES resistant donor plants

[1249] ToLCNDV-ES resistance in wild-grown melon plants was tested according to the tests described under “General Methods.” Wild-grown donor plants (belonging to the subspecies *Melon longifolia* based on origin) exhibiting high resistance to ToLCNDV-ES infection were identified. For resistance, the germplasm was not fixed; therefore, several seed batches were tested to fix and confirm resistance. The resistance level in the fixed germplasm was 9 (as further seen below, the average disease score was 9, while susceptible plants had an average score of approximately 2 to 3). See also Figure 5 See below. qPCR was also performed, showing very little virus in the resistant germline; see [link to qPCR]. Figure 6 And the following text.

[1250] Example 2 - Identification of the genomic location of ToLCNDV resistance

[1251] An F2 mapping population was generated between the resistant donor germplasm and the susceptible culture line. QTL mapping was performed on 200 individuals from the F2 population. A peak marker at 94.2 cM on chromosome 5 was revealed, explaining 99.17% of the variation allocated to a physical location ranging from 25,283,538 Mbp to 26,983,782 Mbp on chromosome 5. This interval was further reduced after fine mapping. Approximately 80 genes were found in this region. Evaluation of the trait's inheritance revealed a single-gene dominance.

[1252] Example 3 - ToLCNDV-ES resistance level

[1253] In the table below and Figure 5 The ToLCNDV-ES symptom levels are presented in the document. Symptom levels were determined at 40 days post-infection (dpi) according to the methods described under “General Methods” in this document.

[1254] average value STDV Piel de Sapo is susceptible 2,3 0,5 VEDRANTAIS is susceptible 2,3 0,5 Galia is susceptible 2,8 0,6 NCIMB 44139 (QTL5rb) 9,0 0,0

[1255] Mean disease score on a 1-9 scale at 40 dpi: mean of the tested genotypes (N=15), STDV: standard deviation; dpi = number of days post-vaccination / infection.

[1256] qPCR testing was also performed. The results are shown below. Figure 6 The figure shows the average q-PCR value (ct value) of 10 plants (N=10) for each genotype at 55 dpi.

[1257] Therefore, at 55 dpi, qPCR (quantitative PCR) was performed on the upper leaves of infected donor plants and susceptible control lines. At 55 dpi, ToLCNDV-ES virus was not detected in the donor tissue, but it was detected in the susceptible controls. (Note that a low Ct value indicates a high viral titer, while a high Ct value indicates a low viral load).

[1258] genotype Average Ct value (55 dpi) Mean standard deviation VEDRANTAIS (Susceptible) 4.1 0.5 NCIMB 44139 (QTL5rb) 30.0 3.0

[1259] The results showed that ToLCNDV-ES virus particles were very low (e.g., average Ct value of at least 30 or higher) or undetectable in the upper leaves of the donor plant NCIMB 44139 after infection.

[1260] Example 4 - SNP markers linked to QTL5rb

[1261] Fine-tuning yielded 7 SNP markers closely linked to QTL5rb.

[1262] Table 2

[1263] Logo Name SNP markers Marker sequence strand orientation relative to the reference genome DHL92 V4 SNP SNP genotype (homozygous) of the resistance donor SNP haplotypes of donor nucleotides containing one or more or all of the SNP markers Location of SNPs on the melon reference genome (DHL92) V4 (melonomics.net database) mME75914 SNP_01 (nucleotide 81 of SEQ ID NO: 1) Forward (+chain) C / T CC C 25456098 mME50729 SNP_02 (nucleotide 242 of SEQ ID NO: 2) Reverse (-chain) C / T GG (or CC when referring to the complementary chain) G (C when referring to the complementary chain) 25849094 mME77064 SNP_03 (nucleotide 81 of SEQ ID NO: 3) Forward (+chain) A / G GG G 25956264 mME72209 SNP_04 (nucleotide 101 of SEQ ID NO: 4) Reverse (-chain) A / G TT (AA when referring to complementary chains) T (A when referring to the complementary chain) 26040072 mME72207 SNP_05 (nucleotide 101 of SEQ ID NO: 5) Reverse (-chain) C / T AA (TT when referring to complementary chains) A (T when referring to the complementary chain) 26142416 mME18971 SNP_06 (nucleotide 101 of SEQ ID NO: 6) Reverse (-chain) A / G TT (AA when referring to complementary chains) T (A when referring to the complementary chain) 26142943 mME75951 SNP_07 (nucleotide 81 of SEQ ID NO: 7) Forward (+chain) C / T CC C 26390451

[1264] It should be noted that the DNA sequences of SNP_02, SNP_04, SNP_05, and SNP_06 are reverse strands (-strands) relative to the reference genome sequence of melon (DHL92) V4 found on the World Wide Web melonomics.net. This can be seen through BLAST analysis of the sequences against the reference genome database, resulting in alignments with the forward strand (where nucleotide numbers are counted upwards) or with the reverse / complementary strand (where nucleotide numbers are counted downwards). It should be understood that this is merely a matter of nomenclature. Thus, nucleotide G of SNP_02 at nucleotide 242 in SEQ ID NO: 2 corresponds to nucleotide C of SNP_02 at nucleotide 242 in the complementary sequence of SEQ ID NO: 2. When referring to donor SNP nucleotides in the reverse strand sequence, such references therefore also include references to complementary donor SNP nucleotides in the complementary / forward strand of the sequence (and vice versa, references to donor nucleotides in the forward strand sequence can also be made by referring to complementary donor nucleotides in the complementary strand).

[1265] The donor nucleotide for any SNP marker described herein may be present in the mentioned SEQ ID NO: or at an equivalent (or corresponding) nucleotide of a sequence having at least 94% (or at least 95%, 96%, 97%, 98%, 99%) sequence identity with a given sequence. This applies to all SNP markers. “Equivalent nucleotides” can be readily identified by pairwise pairing, for example using Emboss-Needle pairwise pairing (using default parameters).

[1266] The donors of QTL5rb and the SNP haplotypes of QTL5 in US11,591,611 are given below. It should be noted that for SNP_02, SNP_04, SNP_05, and SNP_06, the determinations are based on detecting the SNP nucleotide in the reverse strand, which is converted to the SNP nucleotide in the complementary (forward) strand in the data presented below.

[1267] Table 3

[1268] Logo Name SNP haplotype of QTL5rb donor SNP haplotype of QTL5 in US11591611 The name of SNP in US11591611 mME32395 G G SNP_05 in the US patent mME75914 SNP_01 C T mME50729 SNP_02 G A SNP_04 in the US patent mME77064 SNP_03 G A mME72209 SNP_04 T C SNP_08 in the US patent mME72207 SNP_05 A G SNP_07 in the US patent mME18971 SNP_06 T T mME75951 SNP_07 C C mME10621 T T SNP_03 in the US patent mME75966 C C mME43070 G T SNP_02 in the US patent

[1269] It can be seen that the SNP haplotype is different between the marker linked to QTL5rb and QTL5 in US11591611.

[1270] Example 5 - KASP Measurement

[1271] Development of a KASP assay for detecting and / or selecting plants with QTL5rb

[1272] A KASP assay was developed to identify SNPs linked to QTL5rb. SNPs linked to QTL5rb can be identified using the following primers in the KASP assay:

[1273] Common primers for SNPFAM allele and VIC allele

[1274] SNP_01SEQ ID NO 15SEQ ID NO 16SEQ ID NO 17

[1275] SNP_02SEQ ID NO 18SEQ ID NO 19SEQ ID NO 20

[1276] SNP_03SEQ ID NO 21SEQ ID NO 22SEQ ID NO 23

[1277] SNP_04SEQ ID NO 24SEQ ID NO 25SEQ ID NO 26

[1278] SNP_05SEQ ID NO 27SEQ ID NO 28SEQ ID NO 29

[1279] SNP_06SEQ ID NO 30SEQ ID NO 31SEQ ID NO 32

[1280] SNP_07SEQ ID NO 33SEQ ID NO 34SEQ ID NO 35

[1281] Example 6 - Identification of potential causal genes for QTL5rb

[1282] A region ranging from 25,283,538 bp to 26,983,782 bp on chromosome 5 of the melon strain is linked to ToLCNDV resistance observed in the donor. This region contains a large number of TIR-NBS-LRR genes.

[1283] A catalog of TIR-NBS-LRR genes in the QTLrb region was created, resulting in a list of 22 candidate TIR-NBS-LRR genes.

[1284] Next, Illumina short-read sequencing data from wild-type donors were used to identify structural variants in the QTLrb region that were donor-specific (when compared with sequencing data from the melon reference genome and the Vedrantais negative control). This revealed several large deletions co-located or closely adjacent to several candidate TIR-NBS-LRR genes. The largest deletion was 79,597 bp in size and resulted in the deletion of five TIR-NBS-LRR genes. This reduced the list of candidate genes to 17. During the structural variant analysis, it became clear that the QTL5rb region was a highly repetitive region, and therefore the comprehensive gene annotation of the reference genome DHL92(v4) contained many errors in this region. Therefore, the list of the remaining 17 candidate genes was manually compiled using RNAseq data, reducing the number of candidate genes to 7.

[1285] Then, the same Illumina short-read sequencing data was used again to search for donor-specific SNPs or InDels. By applying manually annotated data from the remaining seven TIR-NBS-LRR genes, it was then possible to determine which variants among these variants caused changes in the protein sequence or expression of these genes. This led to the identification of a gene that is a potential causal gene for QTLrb.

[1286] This gene encodes the TIR-NBS-LRR protein (SEQ ID NO: 8). Proteins from the QTL5 donor of US11591611 (SEQ ID NO: 9) were analyzed, and both proteins were compared with those from the reference genome DHL92 v4 (SEQ ID NO: 10) and the susceptible variety Vedrantais. Both DHL92v4 and Vedrantais possess the same protein.

[1287] Figure 1 Multiple sequence alignments are shown for SEQ ID NO: 8 (protein of QTL5rb), SEQ ID NO: 9 (protein of QTL5 of US11591611), and SEQ ID NO: 10 (protein of ToLCNDV-ES susceptible strains DHL92v4 and Vedrantais).

[1288] exist Figure 1 Four amino acid changes were observed, which explain the differences between the QTL5 and QTL5rb proteins in the two susceptible plants and those in the other two plants. These are indicated by black squares and solid lines. These are:

[1289] a) Change 1: Insert two S (serine) residues at the start of the protein.

[1290] b) Change 2: Amino acid V (valine) is changed to M (methionine)

[1291] c) Change 3: Amino acid E (glutamic acid) is changed to K (lysine).

[1292] d) Change 4: Amino acid Y (tyrosine) is changed to histidine H (SEQ ID NO: 8) or aspartic acid D (SEQ ID NO: 9).

[1293] These four changes also Figure 2 As shown in [the document / reference]. And... Figure 4 The image shows potential variations in the genome sequence.

[1294] The last two of these changes are leucine-rich repeat sequences, indicated by black squares and dashed lines.

[1295] Therefore, it is believed that one or more of these four differences (variations 1 to 4) between the “resistance protein” and the “susceptibility protein” are the cause of resistance to ToLCNDV-ES.

[1296] Therefore, the potential resistance protein of QTL5rb preferably contains one or more or all of the following amino acids:

[1297] a) Amino acids numbered 10 to 18 are serine (S).

[1298] b) The amino acid at the equivalent position of amino acid 484 in SEQ ID NO: 8 is methionine (M).

[1299] c) The amino acid at the equivalent position of amino acid 629 in SEQ ID NO: 8 is lysine (K).

[1300] d) The amino acid at the equivalent position of amino acid 679 in SEQ ID NO: 8 is either histidine (H) or aspartic acid (D).

[1301] These four changes also Figure 2 As shown in [the image]. Furthermore, in [the image]... Figure 4 The image shows potential mutations in the genome sequence.

[1302] The gene encoding this protein is expressed in at least the leaves and roots of melon plants.

[1303] like Figure 3 As shown, the gene is located between SNP_03 and SNP_04 on chromosome 5.

[1304] Example 7 - Backcross line with QTL5rb

[1305] The QTL5rb locus was backcrossed into Galia, Honey Dew, Piel de Sapo, Cantaloscope, and Anas backgrounds.

[1306] In Spain, greenhouse trials of various backcross lines were conducted along with suitable controls.

[1307] The experimental design is as follows:

[1308] Sowing date: 13 / 02 / 2024

[1309] Vaccination date: 04 / 03 / 2024

[1310] Implantation date: 11 / 03 / 2024

[1311] Scoring Dates: 01 / 04 / 2024 / 17 / 04 / 2024 / 14 / 05 / 2024

[1312] Plants were grown in peat moss in a greenhouse. Backcross lines were tested using a two-replicated randomized block design (randomized groups at sowing), with five infected plants and one simulated inoculated plant per replicate. For controls, eight replicates were sown in each sowing tray: one resistant control and five susceptible controls.

[1313] As further described above, inoculate with virus-infected tobacco whiteflies (15-25 insects per plant when infected over a 48-hour period). The most aggressive ToLCNDV-ES isolate was used, as mentioned above.

[1314] Genotype of backcross strains Average ToLCNDV-ES score BC2F1 Galia Background 9 BC2F1 Galia Background 9 BC1F3 Honeydew Background 9 BC2F2 Honeydew Background 8.54 BC3F1 Honeydew Background 9 BC2F2 Piel de Sapo background 9 BC3F1 Piel de Sapo background 9 BC3F1 Ananas Background 9 BC2F2 Cantaloupe background 9 BC3F1 Cantaloupe background 9 control strain Honeydew background sensitivity comparison 3.1 Piel de Sapo Background Susceptibility Comparison 3.2 Ananas Background Susceptibility Comparison 4.0 Cantaloupe background sensitivity comparison 2.5 Coliseo F1 (QTL5) 3.0 Coliseo F1 QTL5 donor 7.2

[1315] Except for one plant with a BC2F2 Honeydew background, all plants in the backcross strains had a ToLCNDV-ES score of 9.0.

[1316] Genotyping of backcross lines was also performed based on the SNP markers of the QTL5rb donor.

[1317] All backcross lines possess SNP haplotypes of QTL5rb donors SNPs 03, 04, and 05. However, donor SNP nucleotides are absent in the backcross lines for SNPs 01 and 02, and for SNPs 06 and 07. Therefore, in one respect, it is sufficient for plants containing QTL5rb to contain donor SNP nucleotides of SNPs 03, 04, and / or 05. PCT / RO / 134 form

Claims

1. A cultivated melon plant or a portion thereof, said cultivated melon plant or a portion thereof comprising a recombinant chromosome 5, said recombinant chromosome 5 comprising an introgression fragment comprising a quantitative trait locus (QTL) named QTL5rb between a cytosine of SNP_01 at nucleotide 81 of SEQ ID NO: 1 (nucleotide 25.456.098) corresponding to chromosome 5 of the melon genome and a cytosine of SNP_07 at nucleotide 81 of SEQ ID NO: 7 (nucleotide 26.390.451) corresponding to chromosome 5 of the melon genome, said QTL5rb, when present in homozygous or heterozygous form, confers resistance to New Delhi Tomato Leaf Curl Virus (ToLCNDV-ES) to said melon plant, and said introgression fragment comprising one or more single nucleotide polymorphism (SNP) markers from the following group: - Guanine at nucleotide 81 of SEQ ID NO: 3, or at an equivalent nucleotide in a sequence having at least 95% sequence identity with SEQ ID NO: 3; - Thymine in the adenine or complementary strand of SNP_04 at nucleotide 101 of SEQ ID NO: 4, or at an equivalent nucleotide in a sequence having at least 95% sequence identity with SEQ ID NO: 4; - Adenine in the thymine or complementary strand of SNP_05 at nucleotide 101 of SEQ ID NO: 5, or at an equivalent nucleotide in a sequence having at least 95% sequence identity with SEQ ID NO:

5.

2. The melon plant according to claim 1, wherein the QTL5rb is located between SNP_03 at nucleotide 81 of SEQ ID NO: 3 corresponding to nucleotide 25.956.264 on chromosome 5 of the melon genome and SNP_04 at nucleotide 101 of SEQ ID NO: 4 corresponding to nucleotide 26.040.072 on chromosome 5 of the melon genome.

3. The melon plant according to claim 1 or 2, wherein the melon plant comprises a gene encoding the protein of SEQ ID NO: 8 or a gene encoding a protein having at least 96% sequence identity with SEQ ID NO: 8, said gene being located between SNP_03 at nucleotide 81 of SEQ ID NO: 3 corresponding to nucleotide 25.956.264 on chromosome 5 of the melon genome and SNP_04 at nucleotide 101 of SEQ ID NO: 4 corresponding to nucleotide 26.040.072 on chromosome 5 of the melon genome.

4. The melon plant or a portion thereof according to claim 1, wherein the QTL5rb can be obtained from seeds whose representative sample has been deposited with NCIMB 44139, or from another wild melon donor containing the following SNP marker haplotypes: - Guanine at nucleotide 81 of SEQ ID NO: 3, or at an equivalent nucleotide in a sequence having at least 95% sequence identity with SEQ ID NO: 3; - Thymine in the adenine or complementary strand of SNP_04 at nucleotide 101 of SEQ ID NO: 4, or at an equivalent nucleotide in a sequence having at least 95% sequence identity with SEQ ID NO: 4; - Adenine in the thymine or complementary strand of SNP_05 at nucleotide 101 of SEQ ID NO: 5, or at an equivalent nucleotide in a sequence having at least 95% sequence identity with SEQ ID NO:

5.

5. Seeds, from which the plant according to any one of claims 1 to 4 can grow.

6. The melon plant according to claim 4, wherein the wild melon donor is a subspecies of the melon species.

7. A melon plant or a portion thereof, said melon plant or part thereof comprising chromosome 5 containing a gene encoding a protein of SEQ ID NO: 8 or a gene encoding a protein having at least 96% sequence identity with SEQ ID NO: 8, wherein said gene, when present in homozygous or heterozygous form, confers resistance to New Delhi Tomato Leaf Curl Virus (ToLCNDV-ES) to said melon plant.

8. The plant or plant part according to claim 7, wherein the protein having at least 96% sequence identity with SEQ ID NO: 8 comprises one or more of the following amino acid variations: - Amino acids numbered 10 to 18 are serine (S). - The amino acid at the equivalent position of amino acid 484 in SEQ ID NO: 8 is methionine (M). - The amino acid at the equivalent position of amino acid 629 in SEQ ID NO: 8 is lysine (K). - The amino acid at the equivalent position of amino acid 679 in SEQ ID NO: 8 is either histidine (H) or aspartic acid (D).

9. The plant or plant part according to claim 8, wherein the change in the gene encoding the protein having at least 96% sequence identity with SEQ ID NO: 8 is performed by targeted gene editing of an endogenous gene encoding the protein of SEQ ID NO: 10, or by mutagenesis, such as by radiation or a chemical mutagen.

10. A method for selecting melon plants containing chromosome 5, comprising an introgression fragment comprising a cytosine of SNP_01 located at nucleotide 81 of SEQ ID NO: 1 at nucleotide 25.456.098 of chromosome 5 corresponding to the melon genome, and a cytosine of SNP_07 located at nucleotide 81 of SEQ ID NO: 7 at nucleotide 26.390.451 of chromosome 5 corresponding to the melon genome, the QTL5rb conferring resistance to New Delhi Tomato Leaf Curl Virus (ToLCNDV-ES) to the melon plant when present in homozygous or heterozygous form, the method comprising selecting plants or plant portions containing one or more single nucleotide polymorphism (SNP) markers from the following group: - Guanine at nucleotide 81 of SEQ ID NO: 3, or at an equivalent nucleotide in a sequence having at least 95% sequence identity with SEQ ID NO: 3; - Thymine in the adenine or complementary strand of SNP_04 at nucleotide 101 of SEQ ID NO: 4, or at an equivalent nucleotide in a sequence having at least 95% sequence identity with SEQ ID NO: 4; - Adenine in the thymine or complementary strand of SNP_05 at nucleotide 101 of SEQ ID NO: 5, or at an equivalent nucleotide in a sequence having at least 95% sequence identity with SEQ ID NO:

5.

11. A method for selecting melon plants containing chromosome 5, comprising an introgression fragment comprising a quantitative trait locus (QTL) named QTL5rb between a cytosine of SNP_01 at nucleotide 81 of SEQ ID NO: 1 (nucleotide 25.456.098) corresponding to chromosome 5 of the melon genome and a cytosine of SNP_07 at nucleotide 81 of SEQ ID NO: 7 (nucleotide 26.390.451) corresponding to chromosome 5 of the melon genome, wherein QTL5rb, when present in homozygous or heterozygous form, confers resistance to Tomato Leaf Curl Virus-ES (ToLCNDV-ES) to the melon plant, the method comprising selecting a plant or plant portion comprising a gene encoding a protein of SEQ ID NO: 8 or a gene encoding a protein having at least 96% sequence identity with SEQ ID NO: 8, wherein the gene is located at SEQ ID NO: 25.956.264 (nucleotide 25.956.264) corresponding to chromosome 5 of the melon genome. The SNP_03 at nucleotide 81 of nucleotide 3 is between the SNP_04 at nucleotide 101 of nucleotide 26.040.072 of chromosome 5 of the melon genome.

12. The method of claim 11, wherein the gene encodes a protein having at least 96% sequence identity with SEQ ID NO: 8, and comprises one or more amino acids selected from the following: - Amino acids numbered 10 to 18 are serine (S). - The amino acid at the equivalent position of amino acid 484 in SEQ ID NO: 8 is methionine (M). - The amino acid at the equivalent position of amino acid 629 in SEQ ID NO: 8 is lysine (K). - The amino acid at the equivalent position of amino acid 679 in SEQ ID NO: 8 is either histidine (H) or aspartic acid (D).

13. A method for modifying an endogenous gene encoding the protein of SEQ ID NO: 10 on chromosome 5 of a melon by targeted gene editing or by mutagenesis, such as by radiation or a chemical mutagen, thereby modifying said gene such that the protein encoded having at least 96% sequence identity with SEQ ID NO: 8 comprises one or more of the following amino acid changes: - Amino acids numbered 10 to 18 are serine (S). - The amino acid at the equivalent position of amino acid 484 in SEQ ID NO: 8 is methionine (M). - The amino acid at the equivalent position of amino acid 629 in SEQ ID NO: 8 is lysine (K). - The amino acid at the equivalent position of amino acid 679 in SEQ ID NO: 8 is either histidine (H) or aspartic acid (D).

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