Polymer markers in melon plants and their application in detecting high levels of polymers in melon plants

Abstract

This invention discloses a polymeric marker for melon plants and its application in detecting melon plant height, belonging to the field of nucleotide biotechnology. The technical problem solved by this invention is how to detect melon plant height or select melons with dwarf plant height. The polymeric marker for melon plants disclosed in this invention is the nucleotide at position 90 of SEQ ID No. 1 in the melon genome, which is either G or A. Experiments have shown that the plant height of homozygous melons with the polymeric marker A is lower than that of heterozygous melons with both A and G at the same site, and also lower than that of homozygous melons with G at the same site. This indicates that the polymeric marker for melon plants is correlated with melon plant height and can be used to select dwarf melons.

Description

Technical Field

[0001] This invention belongs to the field of biotechnology nucleotides, specifically relating to polymeric markers for melon plants and their application in detecting high levels of certain substances in melon plants. Background Technology

[0002] melon( Cucumis melo It is a horticultural crop with a long history of cultivation and a wide distribution, and is loved by consumers in various countries.

[0003] In practical breeding and cultivation, plant type is an important trait, and dwarfing is an important phenotype of plant type. First, dwarf plants have a compact plant type and small crown width, which enables efficient high-density planting and increases yield per unit area. Second, dwarf plants have lodging resistance, which is beneficial for mechanized harvesting and facilitates automated and intelligent agricultural production. Therefore, obtaining and identifying dwarf plants has great economic value.

[0004] Developing molecular markers closely linked to dwarfing plant types can improve the efficiency of cultivating dwarf melon plants and promote the development of the melon industry. Summary of the Invention

[0005] The technical problem to be solved by this invention is how to detect the height of melon plants or to select melons with short plant height.

[0006] To solve the above-mentioned technical problems, the present invention first provides the application of substances that detect polymeric markers in melon plants in the detection or auxiliary detection of high levels of polymers in melon plants;

[0007] The polymeric marker of the melon plant is the nucleotide in the melon genome corresponding to position 90 of SEQ ID No. 1 in the sequence listing, which is either G or A.

[0008] In the above applications, the substance used to detect the polymeric markers of melon plants can be a primer pair composed of P1 and P2 or a set of reagents for detecting the polymeric markers of melon plants.

[0009] P1 and P2 are two single-stranded DNA sequences shown in SEQ ID No. 3 and SEQ ID No. 4 of the sequence listing, respectively.

[0010] The complete set of reagents for detecting the polymeric markers in the melon plants includes P1 and P2.

[0011] In the above applications, the kit may also include the restriction endonuclease XhoI.

[0012] Specifically, the substance used to detect the polymeric markers of melon plants can be the primer pair or a reagent kit consisting of the primer pair and XhoI.

[0013] In the above applications, the plant height of homozygous melons whose genome corresponds to nucleotide G at position 90 of SEQ ID No. 1 in the sequence listing is greater than or candidate greater than that of homozygous melons whose genome corresponds to nucleotide A at position 90 of SEQ ID No. 1 in the sequence listing; the plant height of homozygous melons whose genome corresponds to nucleotide G at position 90 of SEQ ID No. 1 in the sequence listing is greater than or candidate greater than that of heterozygous melons whose genome corresponds to both G and A at position 90 of SEQ ID No. 1 in the sequence listing; and the plant height of heterozygous melons whose genome corresponds to both G and A at position 90 of SEQ ID No. 1 in the sequence listing is greater than or candidate greater than that of homozygous melons whose genome corresponds to nucleotide A at position 90 of SEQ ID No. 1 in the sequence listing.

[0014] This invention also provides a method for detecting or assisting in the detection of melon plant height, the method comprising detecting polymeric markers on the melon plant height and determining the melon plant height according to the following method:

[0015] The plant height of homozygous melons whose genome corresponds to nucleotide G at position 90 of SEQ ID No. 1 in the sequence listing is greater than or candidate greater than that of homozygous melons whose genome corresponds to nucleotide A at position 90 of SEQ ID No. 1 in the sequence listing; the plant height of homozygous melons whose genome corresponds to nucleotide G at position 90 of SEQ ID No. 1 in the sequence listing is greater than or candidate greater than that of heterozygous melons whose genome corresponds to both nucleotides G and A at position 90 of SEQ ID No. 1 in the sequence listing; and the plant height of heterozygous melons whose genome corresponds to both nucleotides G and A at position 90 of SEQ ID No. 1 in the sequence listing is greater than or candidate greater than that of homozygous melons whose genome corresponds to nucleotide A at position 90 of SEQ ID No. 1 in the sequence listing.

[0016] In the above method, the detection of the polymeric markers of the melon plants is performed using the substance used to detect the polymeric markers of the melon plants.

[0017] In the above method, detecting the polymeric marker of the melon plant using the substance for detecting the polymeric marker of the melon plant may include: performing PCR amplification on the melon genomic DNA using P1 and P2 to obtain amplification products; sequencing the amplification products to determine the nucleotide corresponding to position 90 of SEQ ID No. 1 in the melon genome; or, digesting the amplification products with the restriction endonuclease XhoI and detecting the size of the digested products. Melons with two DNA fragments in the digested products are homozygous, and the nucleotide corresponding to position 90 of SEQ ID No. 1 in their genome is A; melons with three DNA fragments in the digested products are heterozygous, and the nucleotides corresponding to position 90 of SEQ ID No. 1 in their genome are A and G; melons with one DNA fragment in the digested products are homozygous, and the nucleotide corresponding to position 90 of SEQ ID No. 1 in their genome is G.

[0018] The substance used to detect the polymeric markers of melon plants is also within the scope of protection of this invention.

[0019] The polymeric markers on the melon plants mentioned above are also within the scope of protection of this invention.

[0020] This invention also provides any of the following applications:

[0021] Application of the polymeric markers of melon plants described in X1) in melon breeding;

[0022] X2) Application of the polymeric markers in melon plants for detecting or assisting in the detection of high levels of high-molecular-weight compounds in melon plants;

[0023] X3) The application of the substances used to detect polymeric markers in melon plants in melon breeding;

[0024] X4) The application of the substance used to detect the polymeric markers of melon plants in the preparation of melon breeding products;

[0025] X5) The application of the substance used to detect the polymeric marker of melon plant height in the preparation of products for detecting or assisting in the detection of melon plant height;

[0026] The application of the method for detecting or assisting in detecting melon plant height described in X6) in melon breeding;

[0027] X7) The application of detecting the substance in the melon genome corresponding to nucleotide 90 of SEQ ID No. 1 in the sequence listing in the breeding of melons with dwarf plants and tall stature;

[0028] X8) The application of detecting the substance in the melon genome corresponding to the 90th nucleotide of SEQ ID No. 1 in the sequence listing in the preparation of products with dwarf and tall melons.

[0029] This invention also provides a method for melon breeding, the method comprising Y1) or Y2).

[0030] Y1) Detect the 90th nucleotide in the melon genome corresponding to SEQ ID No.1 in the sequence listing, and select homozygous or heterozygous melons whose 90th nucleotide in the melon genome corresponding to SEQ ID No.1 in the sequence listing is A as parents for breeding;

[0031] Y2) The 90th nucleotide in the melon genome corresponding to SEQ ID No.1 in the sequence listing is mutated from G to A to obtain the target melon, thus realizing melon breeding.

[0032] In one embodiment of the present invention, the melon is a melon ( Cucumis melo The offspring of the S52-8-F2 strain.

[0033] Experiments have shown that the plant height of the aa genotype melon (nucleotide A at position 90 of SEQ ID No. 1) is significantly lower than that of the Aa genotype (nucleotides A and G at position 90 of SEQ ID No. 1) and the AA genotype (nucleotide G at position 90 of SEQ ID No. 1). The plant height of the Aa genotype melon is also significantly lower than that of the AA genotype. This indicates that the molecular marker for melon plant height in this invention is correlated with plant height and can be used to breed dwarf melons. Furthermore, the molecular marker for melon plant height in this invention is correlated with plant height but has little effect on stem diameter and does not affect melon length. This means that the heterozygous type has significant application value in actual production, and can maintain production capacity while reducing production costs.

[0034] The present invention will now be described in further detail with reference to specific embodiments. The given embodiments are merely illustrative of the invention and not intended to limit its scope. The embodiments provided below can serve as a guide for further improvements by those skilled in the art and do not constitute a limitation on the invention in any way.

[0035] In this document, unless otherwise defined herein, terms should be understood according to their common usage by those skilled in the art. Examples of resources describing many of the molecular biology-related terms used herein can be found in the following references: Alberts et al., Molecular Biology of The Cell, 5th ed., Garland Science Publishing, Inc.: New York, 2007; Rieger et al., Glossary of Genetics: Classical and Molecular, 5th ed., Springer-Verlag: New York, 1991; King et al., A Dictionary of Genetics, 6th ed., Oxford University Press: New York, 2002; and Lewin, GenesIX, Oxford University Press: New York, 2007.

[0036] Instructions for the Preservation of Biological Materials

[0037] Abbreviation of depositary institution: CGMCC.

[0038] Name of the depository: China General Microbiological Culture Collection Center, China Microbial Culture Collection Committee.

[0039] Address of the depository: No. 3, No. 1 Courtyard, Beichen West Road, Chaoyang District, Beijing, Postcode: 100101.

[0040] Preservation date: April 21, 2025.

[0041] Registration number at the Preservation Center: CGMCC No. 30706.

[0042] Classification and naming: Melon ( Cucumis melo ).

[0043] Plant number: S52-8-F2. Attached Figure Description

[0044] Figure 1Genetic analysis of the dwarf mutant plant 13C-S52. a. Dwarf plants were obtained through EMS mutagenesis; the white box represents 13C-S52. b. Phenotypic segregation in the F2 population of 13C-S52 conformed to Mendel's 1:2:1 segregation law, meaning the dwarf mutant phenotype was controlled by a recessive single gene. BC1F2 represents the F2 population. c. MutMap pooled sequencing was performed on the F2 population. The SNP index is shown in the figure. Twenty loci with an SNP index of 1 on chromosomes 1, 3, 4, 8, and 9 of the melon were selected for subsequent candidate gene analysis.

[0045] Figure 2 Candidate gene analysis. a, b. Fine mapping to four loci on chromosome 8 of the melon. In b, the left side of the figure shows the melon strain name, 13C represents 13C-WT, S52 represents 13C-S52, and the rest are the names of F2 generation plants. The right side shows the locus information. ce. Phenotype of plants linked at the four loci; c. All four loci are homozygous dominant; d. All four loci are heterozygous; e. All four loci are homozygous recessive. Scale bar = 10cm. f, g. Phenotypes of recombinant single plants; f. heterozygous at chr08_34157772, homozygous dominant at chr08_35184204, chr08_35240383, and chr08_35736165; g. homozygous recessive at chr08_34157772, heterozygous at chr08_35184204, chr08_35240383, and chr08_35736165. Scale bar = 4 cm.

[0046] Figure 3 Fine mapping of the CmTUB8 gene. Among them, BC1F3-46 represents the self-crossed offspring of F2-46 in the F2 generation, BC1F3-118 represents the self-crossed offspring of F2-118 in the F2 generation, A represents homozygous dominant, H represents heterozygous, and B represents homozygous recessive.

[0047] Figure 4Phenotypic and data analysis of plants with genotypes AA, Aa, and aa. ac, scale bar = 10 cm, a. AA genotype plants have normal height, b. Aa genotype plants are semi-dwarfed, c. aa genotype plants are extremely dwarfed. dg, scale bar = 8 cm, each figure from left to right represents AA, Aa, and aa genotype plants, d is a lateral branch at the same node, e is a leaf at the same node, fg is the hermaphroditic and male flowers on the day of flowering. h. Internode length from bottom to top (cotyledonary nodes are not included in the statistics), i. Internode length from bottom to top (8th to 16th nodes), j. Plant height, k. Stem diameter at the 8th node from bottom to top, l. Stem diameter at the 16th node from bottom to top, m. Statistical analysis of the length of hermaphroditic flowers on the day of flowering. In hm, the horizontal axis from left to right represents plants with genotype AA, plants with genotype Aa, and plants with genotype aa, respectively.

[0048] BC1F3-A, BC1F3-H, and BC1F3-B represent plants with genotypes AA, Aa, and aa, respectively.

[0049] In the same bar chart, the same letter indicates no significant difference, while different letters indicate a significant difference (two-way ANOVA).

[0050] Figure 5 Electrophoresis results of enzyme digestion products. A represents the AA genotype, B represents the aa genotype, and H represents the Aa genotype. Detailed Implementation

[0051] Unless otherwise specified, the experimental methods used in the following examples are conventional methods, performed according to the techniques or conditions described in the literature in this field or according to the product instructions. Unless otherwise specified, the materials, reagents, instruments, etc., used in the following examples are commercially available.

[0052] Example 1: The molecular markers for melon plant height are molecular markers related to melon plant height.

[0053] 1. Genetic analysis of the dwarf mutant plant 13C-S52

[0054] The 13C inbred line (13C-WT) is a wild-type, thin-skinned melon variety. 13C-S52 was obtained from the EMS mutation of the 13C-WT melon, representing the M1 generation. Figure 1(a) The M1 generation showed significant phenotypic differences from the wild-type 13C-WT, exhibiting dwarfing, reduced interstitial length, fewer stem nodes, smaller leaves (including leaf width and length), and smaller floral organs. M1 generation was self-crossed to obtain the M2 generation. To remove background noise, the M2 generation was crossed with 13C-WT to obtain the F1 generation. The F1 generation was then self-crossed to obtain the F2 generation. The F2 generation showed phenotypic segregation. Among 160 F2 plants, 55 were of normal height, 75 were of medium height, and 30 were dwarfing mutants, conforming to Mendelian inheritance patterns of 1:2:1 segregation (χ²). 2 =8.4<χ 2 0.01 =9.21), indicating that melon dwarfing is controlled by a recessive single gene ( Figure 1 (b)

[0055] One of the F1 strains is named S52-BC1F1-8, and its self-pollination offspring are designated as the S52-8-F2 strain (i.e., BC1F2). The S52-8-F2 strain was deposited at the China General Microbiological Culture Collection Center on April 21, 2025, with the accession number CGMCC No. 30706.

[0056] 2. Candidate gene analysis and fine mapping of the CmTUB8 gene

[0057] The inventors selected 13 dwarf mutants and 25 normal-height mutants with extreme phenotypic differences from the BC1F2 generation. DNA was extracted from young shoot tips and leaves. The samples were sequenced using the MutMap method and the MGI-DNBSeq platform. The sequencing read length was PE150, and the sequencing data volume was 10 G. Raw reads from the original sequencing sequences were filtered, and the filtered clean reads were further quality controlled and interpreted using the advanced quality control software FastQC, yielding variant SNP information for the W-pool (normal height pool) and M-pool (dwarf mutant pool). For SNPs in both pools, a total of 1195 SNPs with mutations from guanine to adenine or cytosine to thymine were extracted. SNPs with an SNP-index ≤ 0.5 in the W-pool and an SNP-index ≥ 0.3 in the M-pool were retained. Then, using a sliding window of five consecutive SNPs, one SNP was moved at a time to obtain the SNP index regression line. Figure 1 (c)

[0058] The inventors selected 25 SNP-index=1 loci located on chromosomes 1, 3, 4, 8, and 9 of the melon. Primers were designed using Primer Premier 5 for sequencing identification, excluding loci inconsistent with the dwarfing mutant phenotype. Ultimately, the target was narrowed down to four candidate loci, all located on chromosome 8 of the melon. Figure 2 (a). Then, using CAPs and dCAPS molecular markers designed with the dCAPSFinder 2.0 tool (http: / / helix.wustl.edu / dcaps / dcaps.html), four candidate loci were identified: chr08_34157772, chr08_35184204, chr08_35240383, and chr08_35736165. These four loci are often inherited together as dominant, heterozygous, or recessive genotypes, but there is a recombination rate of 4.38%, indicating incomplete linkage. Figure 2 The recombinant single plants exhibited the following characteristics: (1) When the chr08_34157772 locus was dominant homozygous and the chr08_35184204, chr08_35240383, and chr08_35736165 loci were heterozygous, the plant height was normal; (2) When the chr08_34157772 locus was heterozygous and the chr08_35184204, chr08_35240383, and chr08_35736165 loci were recessive homozygous, the plant was semi-dwarfed; (3) When the chr08_34157772 locus was heterozygous and the chr08_35184204, chr08_35240383, and chr08_35736165 locus were dominant homozygous, the plant was semi-dwarfed. Figure 2 (4) When the chr08_34157772 locus is recessive homozygous and the chr08_35184204, chr08_35240383 and chr08_35736165 loci are heterozygous, the plant is dwarfed (f); Figure 2 (g). Here, the inventors found that only the chr08_34157772 locus completely cosegregated with the dwarfing phenotype, and chr08_34157772 was predicted to be a heterosense mutation, chr08_35184204 was in the intergenic region, chr08_35240383 was a synonymous mutation, and chr08_35736165 was a downstream variant. Therefore, the inventors hypothesized that chr08_34157772 was the target locus. Figure 3 Among them, dominant homozygosity means that the locus is homozygous with the same type as the wild type, and recessive homozygosity means that the locus is homozygous with a different type from the wild type (i.e., homozygous after mutation).

[0059] The chr08_34157772 site is located in the coding region of the MELO3C003218 gene, which encodes β-tubulin in melon. This gene is homologous to TUB8 in Arabidopsis thaliana and is named CmTUB8. At the chr08_34157772 site, position 1366 of the CmTUB8 gene (position 422 of the CDS) is mutated from cytosine nucleotide to thymine nucleotide, resulting in a mutation from proline to leucine at position 141 of the amino acid chain.

[0060] Next, to further verify this, the inventors retained the BC1F2 generation recombinant plants BC1F2-46 and BC1F2-118, where chr08_34157772 was heterozygous and chr08_35184204, chr08_35240383, and chr08_35736165 were all homozygous dominant. These were then self-crossed to obtain the BC1F3 generation, which showed three phenotypes: dwarf, intermediate, and normal height. This demonstrates that the chr08_34157772 locus indeed co-segregates with the dwarf phenotype. Figure 4 (ac), and this site was recorded as a polymeric marker for the melon plant.

[0061] 3. Analysis of molecular markers for plant height in BC1F3 generation and melon plant height

[0062] 3.1 Primer Design

[0063] In the 13C-WT genome of melon, the sequences upstream and downstream of the melon plant polymer marker are shown in SEQ ID No. 1. In the 13C-S52 genome, the sequences upstream and downstream of the melon plant polymer marker are shown in SEQ ID No. 2. SEQ ID No. 1 and SEQ ID No. 2 differ at position 90. The complementary nucleotide at position 90 of SEQ ID No. 1 in the 13C-WT genome is C, and the complementary nucleotide at position 90 of SEQ ID No. 2 in the 13C-S52 genome is T. Primers for amplifying the melon plant polymer marker were designed based on the upstream and downstream sequences of the melon plant polymer marker. The primers are as follows:

[0064] P1: 5'-ACATAGCAGAGGCGGTTAAGTAGCG-3' (SEQ ID No. 3);

[0065] P2: 5'-TGACTTGCTGTTTGAGATTCCCGG-3' (SEQ ID No. 4).

[0066] 3.2 Detection of polymeric markers in BC1F3 generation melon plants

[0067] Genomic DNA was extracted from the self-pollinated progeny (BC1F3 generation, a total of 128 plants) of the S52-8-F2 line and amplified by PCR using primers P1 and P2. The PCR reaction mixture consisted of: 7 μL of 2 × Rapid Taq Master Mix (Vazyme), 0.5 μL of primer P1, 0.5 μL of primer P2, 0.5 μL of genomic DNA, and 5.5 μL of ddH2O. The concentration of primers P1 and P2 in the mixture was 10 μM.

[0068] The obtained PCR reaction system was subjected to PCR amplification under the following conditions: 95℃ pre-denaturation for 3 min; 95℃ denaturation for 15 s, 56℃ annealing for 15 s, 72℃ extension for 30 s, 30 cycles, 72℃ extension for 5 min.

[0069] The obtained PCR amplification product was digested with XhoI. The 14 μL digestion reaction system consisted of: 7 μL PCR amplification product, 1.4 μL rcutsmart buffer, 0.2 μL XhoI, and 5.4 μL ddH2O. Both rcutsmart buffer and XhoI were NEB products.

[0070] The enzyme digestion reaction system was digested at 37°C for 6 hours to obtain the enzyme digestion product.

[0071] The enzyme digestion products were then electrophoresed on a 4% agarose gel at 120V for 50 min, and observed and recorded under a gel imaging system after nucleic acid staining with M5 Gelred Plus.

[0072] Melons with two DNA fragments in the enzyme digestion product and two bands on electrophoresis are homozygous. Their genome corresponds to nucleotide A at position 90 of SEQ ID No. 1, designated as the aa genotype melon, with band sizes of 86 and 157 bp. Melons with three DNA fragments in the enzyme digestion product and three bands on electrophoresis are heterozygous. Their genome corresponds to nucleotides A and G at position 90 of SEQ ID No. 1, designated as the Aa genotype melon, with band sizes of 86, 157, and 243 bp. Melons with one DNA fragment in the enzyme digestion product and one band on electrophoresis are homozygous. Their genome corresponds to nucleotide G at position 90 of SEQ ID No. 1, designated as the AA genotype melon, with a band size of 243 bp. Electrophoresis results for some plants are shown below. Figure 5 As shown.

[0073] The results showed that among the 128 plants tested, there were 21 plants with the aa genotype, 65 with the Aa genotype, and 42 with the AA genotype. The aa genotype melons all exhibited extreme dwarfism, the Aa genotype melons all exhibited semi-dwarfism, and the AA genotype melons had normal plant height. The overall height of the 128 plants was measured after 2.5 months of natural climbing. Figure 4 The results (j) show that the average plant height of the AA genotype melon is 212.5 cm, the average plant height of the Aa genotype melon is 139.0 cm, and the average plant height of the aa genotype melon is 31.7 cm. The plant height of the aa genotype melon is significantly lower than that of the Aa and AA genotype melons, and the plant height of the Aa genotype melon is also significantly lower than that of the AA genotype melon. This indicates that the molecular marker for melon plant height in this invention is related to the plant height of melons and can be used to breed dwarf melons.

[0074] 4. Morphological and cytological analysis of CmTUB8 single nucleotide mutant progeny

[0075] Field observations revealed that the CmTUB8 single nucleotide mutation resulted in extreme dwarfing of plants, reduced lateral branch length, decreased number of stem nodes, smaller leaves (including leaf width and length), and smaller floral organs. Figure 4 (ag). Next, the inventors selected 13 AA genotype melon plants, 14 Aa genotype melon plants, and 14 aa genotype melon plants from 128 BC1F3 generation plants that had been planted for two months. The internode lengths from the bottom to the 1st to 8th nodes and from the 8th to 16th nodes, as well as the stem diameters at the 8th and 16th nodes, were measured on these plants after 2.5 months of planting. Figure 4 (h, i, k, l). A two-way ANOVA was performed using GraphPad Prism version 9.5.1 for Windows (GraphPad Software, Massachusetts, Boston, www.graphpad.com), followed by Bonferroni multiple comparison tests. Results showed that the mean internode lengths for the AA, Aa, and aa genotypes were 29.4, 26.1, and 12.7 cm for nodes 1-8, respectively; and 49.8, 38.5, and 14.9 cm for nodes 8-16, respectively. The mean stem diameters for the 8th node were 5.0, 4.9, and 3.5 mm, respectively, and for the 16th node were 5.4, 4.9, and 2.9 mm, respectively. The internode lengths of the aa genotype plants were significantly shorter, while the internode lengths of the Aa genotype plants were between those of the aa and AA genotypes. However, there were significant differences in internode lengths between the AA and aa genotype plants. Figure 4(h,i). The stem diameter of the aa genotype plants was significantly smaller than that of the Aa and AA genotype plants. Similarly, the stem diameter of the Aa genotype was between that of the aa and AA genotypes, but there was no significant difference in stem diameter between the Aa and AA genotype plants in Section 8. Figure 4 (k,l).

[0076] In addition, the inventors selected 16 hermaphroditic flowers from AA genotype plants, 13 from Aa genotype plants, and 13 from aa genotype plants on the day of flowering, and measured the length of the hermaphroditic flowers (i.e., the length from the base of the pedicel to the base of the petals) for each genotype. They found that the average lengths of the hermaphroditic flowers from the AA, Aa, and aa genotypes were 27.0, 25.1, and 18.5 mm, respectively. The hermaphroditic flowers from the aa genotype plants were significantly smaller, while there was no significant difference in length between the hermaphroditic flowers from the Aa and AA genotype plants. Figure 4 (m).

[0077] The results indicate that the C-to-T heterozygous mutation at the chr08_34157772 site of the CmTUB8 gene only affects plant height, has a smaller impact on stem diameter, and does not affect fruit length. This means that the heterozygote has great application value in actual production, reducing production costs while maintaining production capacity.

[0078] The present invention has been described in detail above. For those skilled in the art, the invention can be practiced in a wide range of ways with equivalent parameters, concentrations, and conditions without departing from its spirit and scope, and without requiring unnecessary experiments. Although specific embodiments have been given, it should be understood that further modifications can be made to the invention. In summary, according to the principles of the invention, this application is intended to include any changes, uses, or improvements to the invention, including changes made using conventional techniques known in the art that depart from the scope disclosed herein. Some of the essential features can be applied within the scope of the following appended claims.

Claims

1. Application of substances containing polymeric markers in the detection or auxiliary detection of high levels of certain substances in melon plants; The polymeric marker of the melon plant is the nucleotide in the melon genome corresponding to position 90 of SEQ ID No. 1 in the sequence listing, which is either G or A.

2. The application according to claim 1, characterized in that: The substance used to detect the polymeric markers in melon plants is a primer pair composed of P1 and P2 or a set of reagents for detecting the polymeric markers in melon plants. P1 and P2 are two single-stranded DNA sequences shown in SEQ ID No. 3 and SEQ ID No. 4 of the sequence listing, respectively. The complete set of reagents for detecting the polymeric markers in the melon plants includes P1 and P2.

3. The application according to claim 2, characterized in that: The kit also includes the restriction endonuclease XhoI.

4. The application according to any one of claims 1-3, characterized in that: The plant height of homozygous melons whose nucleotide at position 90 of SEQ ID No. 1 in the genome is G is greater than or candidate greater than the plant height of homozygous melons whose nucleotide at position 90 of SEQ ID No. 1 in the genome is A; the plant height of homozygous melons whose nucleotide at position 90 of SEQ ID No. 1 in the genome is G or candidate greater than the plant height of heterozygous melons whose nucleotides at positions 90 of SEQ ID No. 1 in the genome are both G and A; and the plant height of heterozygous melons whose nucleotides at positions 90 of SEQ ID No. 1 in the genome are both G and A.

5. A method for detecting or assisting in the detection of melon plant height, characterized in that: The method includes detecting the polymeric markers of the melon plant as described in claim 1, and determining the height of the melon plant according to the following method: The plant height of homozygous melons whose genome corresponds to nucleotide G at position 90 of SEQ ID No. 1 in the sequence listing is greater than or candidate greater than that of homozygous melons whose genome corresponds to nucleotide A at position 90 of SEQ ID No. 1 in the sequence listing; the plant height of homozygous melons whose genome corresponds to nucleotide G at position 90 of SEQ ID No. 1 in the sequence listing is greater than or candidate greater than that of heterozygous melons whose genome corresponds to both nucleotides G and A at position 90 of SEQ ID No. 1 in the sequence listing; and the plant height of heterozygous melons whose genome corresponds to both nucleotides G and A at position 90 of SEQ ID No. 1 in the sequence listing is greater than or candidate greater than that of homozygous melons whose genome corresponds to nucleotide A at position 90 of SEQ ID No. 1 in the sequence listing.

6. The method according to claim 5, characterized in that: The detection of the melon plant polymer markers described in claim 1 is performed using any of the substances described in claims 1-3 for detecting melon plant polymer markers.

7. Any of the following applications: X1) The application of the polymeric markers for melon plants as described in claim 1 in melon breeding; X2) The application of the polymeric markers for melon plants as described in claim 1 in the detection or auxiliary detection of high levels of polymers in melon plants; X3) The application of any of the substances described in claims 1-3 for detecting polymeric markers in melon plants in melon breeding; X4) The use of any of the substances described in claims 1-3 for detecting the polymeric markers of melon plants in the preparation of products for detecting or assisting in the detection of melon plant height; X5) The application of the method according to claim 5 or 6 in melon breeding; X6) The application of detecting the substance in the melon genome corresponding to the 90th nucleotide of SEQ ID No. 1 in the sequence listing in the breeding of melons with dwarf plants and tall stature.

8. Melon breeding methods, including: The 90th nucleotide in the melon genome corresponding to SEQ ID No. 1 in the sequence listing was detected, and homozygous or heterozygous melons whose 90th nucleotide in the melon genome corresponding to SEQ ID No. 1 in the sequence listing was A were selected as parents for breeding.

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