SNP (Single Nucleotide Polymorphism) molecular marker related to insect resistance of sugarcane and application of SNP molecular marker

Abstract

The invention belongs to the technical field of plant genetic engineering, and provides an SNP (Single Nucleotide Polymorphism) molecular marker related to insect resistance of sugarcane and application of the SNP molecular marker. The method comprises the following steps: screening to obtain an insect-resistant variety Cinnamomum subaequalis and an insect-sensitive variety Cinnamomum subaequalis 57, respectively collecting 30 samples of the two varieties, carrying out whole genome re-sequencing, and carrying out whole genome association analysis to obtain 10 SNP loci located in sugarcane chromosomes Chr1F: 20752, Chr2B: 20889, Chr3D: 384, Chr4J: 17220, Chr5D: 15964, Chr6K: 15569, Chr7D: 3376, Chr8A: 19687, Chr9H: 16936 and Chr10L: 4675, and the coordinates of the ten genomes are strongest in association with the insect-resistant character of sugarcane. The SNP site developed by the invention can be used as an important molecular target for sugarcane insect resistance, and a molecular foundation is laid for creating sugarcane insect-resistant varieties.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of plant genetic engineering, and particularly relates to a SNP molecular marker related to sugarcane insect resistance and application thereof. BACKGROUND

[0002] Sugarcane is an important economic crop, sugar crop and renewable energy plant in China. China is the third largest sugarcane producing country in the world. However, sugarcane production is threatened by various diseases and pests, among which the Lepidoptera boring moth is the most important pest that harms sugarcane. The whole process of sugarcane from planting to harvesting can be harmed by the sugarcane borer. After being harmed in the seedling stage, the seedling cannot germinate and emerge; after being harmed in the seedling stage, the growth point dies to form a dry heart, reducing the number of seedlings and effective stems; in the middle and late growth stages, the cane stem or stem base is bored and eaten, the internal tissue is damaged, the growth of sugarcane is affected, and the sugar content is reduced. When encountering strong winds, the cane stem is often broken at the insect mouth; and the wound left after being eaten by insects is also the entry point of pathogenic fungi such as pink rot disease, causing the overlap of insect pests and diseases, resulting in yield reduction and affecting the ratooning of sugarcane, which poses a serious threat to sugarcane. At present, the prevention and control of sugarcane pests mainly relies on chemical control. However, long-term and large-scale use of various chemical pesticides increases the resistance of pests, gradually reduces the control efficiency, seriously pollutes the environment, and threatens human health and destroys the balance of agricultural ecology.

[0003] Using plant insect-resistant transgenic technology to create sugarcane insect-resistant varieties is one of the most effective measures for the comprehensive control of sugarcane pests. At present, the use of Bt gene is the most widely used in the research of insect-resistant transgenic sugarcane. The protein expressed by Bt gene can specifically kill Lepidoptera, Homoptera and Diptera pests, and is harmless to humans and animals. However, with the wide use of Bt crops, insects have evolved resistance to Bt toxins.

[0004] Therefore, developing new insect-resistant molecular mechanisms, exploring the defense mechanism of sugarcane, and developing SNP molecular markers related to sugarcane insect resistance will fill the gap in the scientific defense mechanism of sugarcane against pests, and provide key targets for the subsequent creation of high-quality insect-resistant germplasm. SUMMARY

[0005] In view of the deficiencies of the prior art, the purpose of the present application is to provide a SNP molecular marker related to the insect resistance of sugarcane and an application thereof. The present application adopts 19 sugarcane varieties to carry out two laboratory insect resistance screening experiments, i.e. sugarcane borer feeding different sugarcane variety leaf area statistics experiment and sugarcane borer feeding different sugarcane variety leaf mortality experiment, and screens out the insect-resistant variety Zheshemi and the insect-susceptible variety Guitang 57. 30 samples of each of the two varieties are collected for whole genome resequencing, and through whole genome association analysis, 10 SNP loci located on the sugarcane chromosome are obtained, and the ten genomic coordinates have the strongest correlation with the insect resistance traits of sugarcane. The SNP loci developed by the present application can be used as an important molecular target for insect resistance of sugarcane, and lay a molecular foundation for creating new germplasm of insect-resistant sugarcane.

[0006] In order to achieve the purpose of the present application, the following technical solutions are adopted:

[0007] In the first aspect, the present application provides a SNP molecular marker related to the insect resistance of sugarcane, wherein the SNP molecular marker is selected from at least one of molecular markers SNP1-SNP10.

[0008] SNP1 is located at sugarcane chromosome Chr1F:20752, and the allele of the SNP locus is C / A;

[0009] SNP2 is located at sugarcane chromosome Chr2B:20889, and the allele of the SNP locus is A / C;

[0010] SNP3 is located at sugarcane chromosome Chr3D:384, and the allele of the SNP locus is A / G;

[0011] SNP4 is located at sugarcane chromosome Chr4J:17220, and the allele of the SNP locus is G / T;

[0012] SNP5 is located at sugarcane chromosome Chr5D:15964, and the allele of the SNP locus is A / T;

[0013] SNP6 is located at sugarcane chromosome Chr6K:15569, and the allele of the SNP locus is A / C;

[0014] SNP7 is located at sugarcane chromosome Chr7D:3376, and the allele of the SNP locus is T / C;

[0015] SNP8 is located at sugarcane chromosome Chr8A:19687, and the allele of the SNP locus is G / A;

[0016] SNP9 is located at sugarcane chromosome Chr9H:16936, and the allele of the SNP locus is G / A;

[0017] The SNP10 is located at sugarcane chromosome Chr10L:4675, and the alleles of the SNP site are T / C.

[0018] In the present application, the alleles of the SNP1 site are C / A; wherein the reference base of the SNP1 site is C, and the variant base is A; when the base of the SNP1 site is A, the corresponding sugarcane has insect resistance. The reference base and the variant base of the other sites are described with reference to the SNP1 site.

[0019] In the second aspect, the present application provides the application of the SNP marker related to the insect resistance of sugarcane in the improvement of the insect-resistant varieties or lines of sugarcane.

[0020] In the present application, all genes within a specific range upstream and downstream of each SNP are found by using the physical position of the SNP marker site related to the insect resistance of sugarcane in the first aspect on the reference genome of sugarcane, the candidate genes related to the plant insect defense mechanism are screened by performing functional annotation analysis on these genes, and the insect-resistant varieties or lines of sugarcane are improved by using these genes.

[0021] In the third aspect, the present application provides the application of the SNP marker related to the insect resistance of sugarcane in the assisted breeding of the insect-resistant varieties of sugarcane.

[0022] In the present application, the assisted breeding of the insect-resistant varieties of sugarcane includes the following steps: extracting the genomic DNA of the sugarcane varieties to be bred, detecting the genotypes of the above-mentioned SNP sites, and selecting the sugarcane varieties with excellent genotypes for breeding according to the correlation between the genotypes and the insect resistance.

[0023] In the fourth aspect, the present application provides a method for detecting the strength of the insect resistance of sugarcane, which comprises the following steps: extracting nucleic acid from a sugarcane sample, detecting the nucleic acid region containing the SNP molecular marker in the first aspect, and judging the strength of the insect resistance of the sugarcane according to the genotype of the detected SNP site.

[0024] Preferably, the detection method comprises direct sequencing, specific probe hybridization, specific primer extension or PCR.

[0025] Preferably, the standard for judging the insect resistance of the sugarcane is that the more alleles of the specific SNP sites are detected, the stronger the insect resistance of the sugarcane variety is.

[0026] The alleles of the SNP sites include: the alleles of the Chr1F:20752 site are A, the alleles of the Chr2B:20889 site are C, the alleles of the Chr3D:384 site are G, the alleles of the Chr4J:17220 site are T, the alleles of the Chr5D:15964 site are T, the alleles of the Chr6K:15569 site are C, the alleles of the Chr7D:3376 site are C, the alleles of the Chr8A:19687 site are A, the alleles of the Chr9H:16936 site are A, and the alleles of the Chr10L:4675 site are C.

[0027] In a fifth aspect, the present application provides an isolated nucleic acid molecule, which comprises a nucleotide sequence as shown in SEQ ID NO:1 or a complementary sequence thereof, wherein the 51th position of the sequence is a SNP site Chr1F:20752, and the alleles of the SNP site are C / A;

[0028] Or, the nucleic acid molecule comprises a nucleotide sequence as shown in SEQ ID NO:2 or a complementary sequence thereof, wherein the 51th position of the sequence is a SNP site Chr2B:20889, and the alleles of the SNP site are A / C;

[0029] Or, the nucleic acid molecule comprises a nucleotide sequence as shown in SEQ ID NO:3 or a complementary sequence thereof, wherein the 51th position of the sequence is a SNP site Chr3D:384, and the alleles of the SNP site are A / G;

[0030] Or, the nucleic acid molecule comprises a nucleotide sequence as shown in SEQ ID NO:4 or a complementary sequence thereof, wherein the 51th position of the sequence is a SNP site Chr4J:17220, and the alleles of the SNP site are G / T;

[0031] Or, the nucleic acid molecule comprises a nucleotide sequence as shown in SEQ ID NO:5 or a complementary sequence thereof, wherein the 51th position of the sequence is a SNP site Chr5D:15964, and the alleles of the SNP site are A / T;

[0032] Or, the nucleic acid molecule comprises a nucleotide sequence as shown in SEQ ID NO:6 or a complementary sequence thereof, wherein the 51th position of the sequence is a SNP site Chr6K:15569, and the alleles of the SNP site are A / C;

[0033] Or, the nucleic acid molecule comprises a nucleotide sequence as set forth in SEQ ID NO: 7 or a complementary sequence thereof, wherein the 51st position of the sequence is SNP site Chr7D:3376, and the allele of the SNP site is T / C;

[0034] Or, the nucleic acid molecule comprises a nucleotide sequence as set forth in SEQ ID NO: 8 or a complementary sequence thereof, wherein the 51st position of the sequence is SNP site Chr8A:19687, and the allele of the SNP site is G / A;

[0035] Or, the nucleic acid molecule comprises a nucleotide sequence as set forth in SEQ ID NO: 9 or a complementary sequence thereof, wherein the 51st position of the sequence is SNP site Chr9H:16936, and the allele of the SNP site is G / A;

[0036] Or, the nucleic acid molecule comprises a nucleotide sequence as set forth in SEQ ID NO: 10 or a complementary sequence thereof, wherein the 51st position of the sequence is SNP site Chr10L:4675, and the allele of the SNP site is T / C.

[0037] In a sixth aspect, the present application provides a primer or probe for detecting the insect resistance of sugarcane, wherein the target of the primer or probe is the isolated nucleic acid molecule of the sixth aspect.

[0038] In a seventh aspect, the present application provides a kit for detecting the insect resistance of sugarcane, wherein the kit comprises: a primer or probe for detecting the SNP site of the first aspect, or the primer or probe for detecting the insect resistance of sugarcane of the sixth aspect.

[0039] In an eighth aspect, the present application provides a SNP typing chip, wherein a specific probe for the SNP site of the first aspect is fixed on a solid carrier of the chip.

[0040] Compared with the prior art, the present application has the following beneficial effects:

[0041] (1) The research results of the present application show that the ten SNP loci located at sugarcane chromosomes Chr1F:20752, Chr2B:20889, Chr3D:384, Chr4J:17220, Chr5D:15964, Chr6K:15569, Chr7D:3376, Chr8A:19687, Chr9H:16936, Chr10L:4675 have the strongest correlation with the insect resistance trait of sugarcane. Using the physical positions of the ten SNP loci on the reference genome of sugarcane, all genes within a specific range upstream and downstream of each SNP are searched, and the candidate genes related to the plant insect resistance defense mechanism are screened through functional annotation analysis, which can provide key targets for subsequent creation of high-quality insect-resistant germplasm.

[0042] (2) The research results of the present application show that the ten SNP marker loci discovered are used to verify the correlation and prediction ability of the SNPs with the insect resistance phenotype in a wider range of sugarcane germplasm resources (especially breeding parents and offspring populations), which can provide key targets for the development of stable, low-cost, high-throughput detection methods and the selection of plants with excellent insect resistance alleles.

[0043] (3) The research results of the present application show that the ten SNP marker loci discovered are used for in-depth study of the strong candidate genes located, and through expression analysis and functional verification, it is revealed how these SNPs affect gene function or expression, and ultimately lead to differences in insect resistance phenotype, which can provide key targets for mining specific defense pathways of sugarcane insect resistance and analyzing the molecular mechanism of sugarcane insect resistance.

[0044] (4) The research results of the present application show that the ten SNP marker loci discovered can provide key targets for evaluating germplasm resources and variety protection, and gene editing and other modern biological technology breeding. BRIEF DESCRIPTION OF DRAWINGS

[0045] Figure 1 is the leaf area of sugarcane leaf eaten by sugarcane borer after 72h of feeding.

[0046] Figure 2 is the statistical result of leaf area of sugarcane leaf eaten by sugarcane borer.

[0047] Figure 3 is a schematic diagram of the experimental method for the mortality of sugarcane borer caused by feeding on different varieties of sugarcane leaves.

[0048] Figure 4 is the survival curve of sugarcane borer feeding on different varieties of sugarcane leaves.

[0049] Figure 5 is the Manhattan plot of the whole genome association analysis result. DETAILED DESCRIPTION

[0050] The technical solutions of the present application are further illustrated below through specific embodiments. Those skilled in the art should understand that the embodiments are only used to help understand the present application and should not be regarded as specific limitations of the present application.

[0051] Unless otherwise specified in the embodiments, the techniques or conditions are carried out according to the techniques or conditions described in the literature in the art or according to the product instructions. Unless otherwise specified, the reagents or instruments used are conventional products that can be commercially available through regular channels.

[0052] The following terms are involved in the specific embodiments:

[0053] Clean Reads: The result of raw reads after quality filtering and error correction.

[0054] SNP: Single nucleotide polymorphism (SNP) mainly refers to the DNA sequence polymorphism caused by a single nucleotide variation at the genomic level of an individual. They are the most abundant DNA molecular marker type in the genome.

[0055] Small InDel: Small insertion or deletion (small insertion / deletion) mainly refers to the insertion or deletion mutation of short fragments (usually 1-50 bp) in the genome, which belongs to a kind of structural genetic variation, can significantly affect gene function through frameshift mutation or regulatory modification, and has basic research value and application potential (such as molecular marker development, gene editing target design).

[0056] Example 1

[0057] Statistical experiment of leaf area of sugarcane borer feeding on different varieties of sugarcane leaves

[0058] 1. Embodiment

[0059] (1) In 2024, the following 19 sugarcane varieties were collected in Nanning, Guangxi University, Guangxi Zhuang Autonomous Region, China: Shinriku 16, Guitang 21, ROC22, Guitang 29, Guitang 42, Guitang 44, Guitang 57, Guitang 58, Guitang 60, Guitang 63, Guitang 67, Guitang 68, Guitang 76, Taisaku 134, Guiliu 05136, Yutang 71 / 210, Yutang 93 / 159, Banmao and Geishami.

[0060] (2) Take sugarcane leaves of different varieties but uniform area (12.5 cm 2), and placed in 90 mm culture dishes with filter paper moistened with water on the bottom of the dish to prevent the sugarcane leaves from wilting during the experiment. Three 3rd instar larvae of A. interrupta were placed on each sugarcane leaf in each dish. Six biological replicates were performed for each variety, and the area of the leaf eaten by the larvae was observed and recorded at 24 h, 48 h, and 72 h after the larvae were allowed to feed. The final feeding results are shown in Table 1. Figure 1

[0061] 2. Results

[0062] The area of the leaf eaten by the larvae was observed and recorded at 24 h, 48 h, and 72 h after the larvae were allowed to feed. The final feeding results are shown in Table 1.

[0063] The results show that the variety GT57 had the largest area of leaf eaten by the larvae, and the variety QSM had the smallest area of leaf eaten by the larvae (Table 1). GraphPad software was used to plot the results and perform statistical multiple comparisons, and the results show that GT57 had a significant difference from NT16, GT42, GT58, and QSM (P < 0.05). The variety GT57 was screened as a susceptible variety, and the varieties NT16, GT42, GT58, and QSM were screened as resistant varieties. Figure 2

[0064] Table 1 shows the results of the area of the leaf eaten by the larvae of A. interrupta on different varieties of sugarcane.

[0065] Table 1

[0066]

[0067] Example 2

[0068] Experiment on the mortality of A. interrupta caused by feeding on leaves of different varieties of sugarcane

[0069] 1. Embodiment

[0070] (1) Based on the screening results obtained in Example 1, the resistant varieties NT16, GT58, and QSM were selected, and the susceptible varieties GT57, GT60, and GT21 were selected.

[0071] (2) The leaves of the six varieties of sugarcane were collected and the veins were removed, as shown in Table 1. The leaves were cut to the size of the diameter of the 24-well plate, and two 24-well plates were filled with leaves of each variety, with one 2nd instar larva placed in each well, for a total of 48 larvae. To maintain a fresh leaf environment for the larvae, the leaves were replaced every two days, and the growth and survival of the larvae were observed and recorded daily. The 48 larvae of each variety were observed and recorded for 13 days. Figure 3

[0072] 2. Results

[0073] The observation and recording results are shown in Table 2.

[0074] ​​​The survival curves of the sugarcane borer were drawn using GraphPad software, and statistical multiple comparisons showed that there was no difference among Guitang 57, Guitang 60 and Guitang 21; Guitang 58 was not different from Guitang 57, but was different from Guitang 60 and Guitang 21; Xintaitang 16 was not different from Jieshemi, but both were different from the others. Figure 4 The susceptible varieties were Guitang 57 and Guitang 60, and the resistant varieties were Xintaitang 16 and Jieshemi.

[0075] In summary, the susceptible variety Guitang 57 and the resistant variety Jieshemi can be selected for subsequent genome resequencing.

[0076] Table 2

[0077]

[0078]

[0079] Example 3

[0080] Development of SNP molecular markers related to sugarcane resistance

[0081] 1. Embodiment

[0082] (1) In 2024, 30 samples of Guitang 57 and Jieshemi were collected at Guangxi University in Nanning, Guangxi Zhuang Autonomous Region, China, and each sample was collected from a single individual. The 60 samples were subjected to GWAS whole genome association analysis.

[0083] (2) After the sample genomic DNA was detected to be qualified, the DNA was fragmented by mechanical disruption (ultrasonic wave), and then the fragmented DNA was subjected to fragment purification, end repair, 3' end A addition, ligation of sequencing adapters, and then subjected to fragment size selection by agarose gel electrophoresis. The sequencing library was formed by PCR amplification, and the library was first subjected to library quality inspection. The qualified library was sequenced by Illumina. The raw reads (double-end sequences) obtained by sequencing were subjected to quality assessment and filtration to obtain Clean Reads, which were used for subsequent bioinformatics analysis. The Clean Reads were aligned with the reference genome sequence, and based on the alignment results, SNP and Small InDel detection were performed. Further whole genome association analysis was performed.

[0084] 2. Results

[0085] (1) Whole genome association analysis

[0086] Trait association analysis:

[0087] Figure 5Figure 2 is a Manhattan plot, which is a plot of the effect size of genetic markers, i.e. genome-wide P values after F test, on the physical position on the chromosome, the horizontal axis is the genome coordinate, and the vertical axis is -log10P, the smaller the P value, the stronger the correlation, which is represented by the larger vertical axis.

[0088] By searching the peak area in the Manhattan plot, it is found that the ten genome coordinates Chr1F:20752, Chr2B:20889, Chr3D:384, Chr4J:17220, Chr5D:15964, Chr6K:15569, Chr7D:3376, Chr8A:19687, Chr9H:16936, and Chr10L:4675 have the strongest correlation with the insect resistance trait.

[0089] The sequences of 50 bp upstream and downstream of the SNP site Chr1F:20752 are shown in SEQ ID NO: 1. The sequences of 50 bp upstream and downstream of the SNP site Chr2B:20889 are shown in SEQ ID NO: 2. The sequences of 50 bp upstream and downstream of the SNP site Chr3D:384 are shown in SEQ ID NO: 3. The sequences of 50 bp upstream and downstream of the SNP site Chr4J:17220 are shown in SEQ ID NO: 4. The sequences of 50 bp upstream and downstream of the SNP site Chr5D:15964 are shown in SEQ ID NO: 5. The sequences of 50 bp upstream and downstream of the SNP site Chr6K:15569 are shown in SEQ ID NO: 6. The sequences of 50 bp upstream and downstream of the SNP site Chr7D:3376 are shown in SEQ ID NO: 7. The sequences of 50 bp upstream and downstream of the SNP site Chr8A:19687 are shown in SEQ ID NO: 8. The sequences of 50 bp upstream and downstream of the SNP site Chr9H:16936 are shown in SEQ ID NO: 9. The sequences of 50 bp upstream and downstream of the SNP site Chr10L:4675 are shown in SEQ ID NO: 10.

[0090] Multiple hypothesis testing correction:

[0091] Multiple hypothesis testing correction can reduce the expansion of type I error and false positive correlation, the number of multiple hypothesis testing depends on the number of SNPs contained in the genome, and we use FDR correction method to correct the P value after multiple hypothesis testing in GWAS analysis to reduce the probability of false positive. The correction results are shown in Table 3, and the FDR of all significant SNP sites is <0.05. As can be seen from Table 3, the results of the ten SNP sites obtained have high reliability, the false positive rate is ≤5%, and it is more likely to reflect the true biological effect rather than random fluctuation, thereby providing high-quality targets for downstream experiments.

[0092] Table 3

[0093] chr rs ps n_mis n_obs allele1 allele0 af beta se p_wald fdr Chr3D Chr3D:384 384 0 60 G A 0.492 -0.50014 0.008477 1.75E-53 6.85E-51 Chr7D Chr7D:3376 3376 0 60 C T 0.492 -0.50014 0.008477 1.75E-53 6.85E-51 Chr10L Chr10L:4675 4675 0 60 C T 0.492 0.500141 0.008477 1.75E-53 6.85E-51 Chr6K Chr6K:15569 15569 0 60 C A 0.492 -0.50014 0.008477 1.75E-53 6.85E-51 Chr5D Chr5D:15964 15964 0 60 T A 0.492 -0.50014 0.008477 1.75E-53 6.85E-51 Chr4J Chr4J:17220 17220 0 60 T G 0.492 -0.50014 0.008477 1.75E-53 6.85E-51 Chr9H Chr9H:16936 16936 0 60 A G 0.483 -0.50058 0.011993 5.77E-45 1.64E-42 Chr1F Chr1F:20752 20752 0 60 A C 0.258 -0.91266 0.029441 8.57E-38 2.14E-36 Chr8A Chr8A:19687 19687 0 60 A G 0.242 -0.76383 0.059702 1.57E-18 1.35E-17 Chr2B Chr2B:20889 20889 0 60 C A 0.15 0.616438 0.105651 2.55E-07 9.76E-07

[0094] Note, in Table 3, chr represents the chromosome number where the SNP is located; rs represents the SNP identifier in dbSNP database; ps represents the physical position of the SNP on the chromosome (unit: bp); n mis represents the number of samples missing the SNP typing (data quality control key indicator); n obs represents the number of samples successfully typed (actual sample size participating in analysis); allele1 represents the effect allele (allele used to calculate the effect in association analysis, usually ALT); allele0 represents the reference allele (used as a comparison benchmark, usually REF); af represents the frequency of the effect allele (range 0-1); beta represents the effect value (the magnitude of the effect of allele1 on the phenotype in linear regression); se represents the standard error of the effect value (measures the accuracy of the beta estimate, the smaller the more reliable); p wald represents the original P value without correction (significance based on Wald test); fdr represents the multiple hypothesis test P value after correction (multiple test correction result, fdr < 0.05 is significant).

[0095] In summary, the present application shows that ten genomic coordinates are most strongly associated with the insect resistance trait of sugarcane by whole genome association analysis, and the above coordinate sites can be used as important molecular targets for insect resistance of sugarcane, and lay a molecular foundation for creating new germplasm of insect-resistant sugarcane.

[0096] The applicant states that the above is only a specific embodiment of the present application, but the protection scope of the present application is not limited thereto. It should be understood by those skilled in the art that any changes or replacements within the technical scope disclosed by the present application can be easily thought of by any person skilled in the art, and all fall within the protection scope and disclosure scope of the present application.

Claims

1. A SNP molecular marker associated with insect resistance in sugarcane, characterized in that: The SNP molecular marker is selected from at least one of the molecular markers SNP1-SNP10; SNP1 is located on sugarcane chromosome Chr1F:20752, and the allele of the SNP site is C / A; SNP2 is located on sugarcane chromosome Chr2B:20889, and the alleles of the SNP site are A / C; SNP3 is located on sugarcane chromosome Chr3D:384, and the alleles of the SNP site are A / G; SNP4 is located on sugarcane chromosome Chr4J:17220, and the allele of the SNP site is G / T; SNP5 is located on sugarcane chromosome Chr5D:15964, and the alleles of the SNP site are A / T; SNP6 is located on sugarcane chromosome Chr6K:15569, and the alleles of the SNP site are A / C; SNP7 is located on sugarcane chromosome Chr7D:3376, and the allele of the SNP site is T / C; SNP8 is located on sugarcane chromosome Chr8A:19687, and the allele of the SNP site is G / A; SNP9 is located on sugarcane chromosome Chr9H:16936, and the allele of the SNP site is G / A; SNP10 is located on sugarcane chromosome Chr10L:4675, and the allele of the SNP site is T / C.

2. Use of the SNP marker associated with insect resistance in sugarcane according to claim 1 in improving insect-resistant sugarcane varieties or lines.

3. Use of the SNP marker associated with insect resistance in sugarcane according to claim 1 in assisting breeding of insect-resistant sugarcane varieties.

4. A method for detecting the insect resistance of sugarcane, characterized in that: The method comprises: extracting nucleic acid from a sugarcane sample, detecting a nucleic acid region containing the SNP molecular marker according to claim 1, and judging the insect resistance of the sugarcane according to the alleles of the detected SNP sites.

5. The method for detecting insect resistance of sugarcane according to claim 4, characterized in that: The detection method includes: direct sequencing, specific probe hybridization method, specific primer extension method or PCR method.

6. The method for detecting insect resistance of sugarcane according to claim 4 or 5, characterized in that: The standard for judging the insect resistance of sugarcane is: the alleles of the SNP locus according to claim 1 are detected, and the more alleles of the specific SNP locus are detected, the stronger the insect resistance of the sugarcane variety is; The alleles of the SNP sites include: the allele at Chr1F:20752 is A, the allele at Chr2B:20889 is C, the allele at Chr3D:384 is G, the allele at Chr4J:17220 is T, the allele at Chr5D:15964 is T, the allele at Chr6K:15569 is C, the allele at Chr7D:3376 is C, the allele at Chr8A:19687 is A, the allele at Chr9H:16936 is A, and the allele at Chr10L:4675 is C.

7. An isolated nucleic acid molecule, characterized in that The nucleic acid molecule comprises a nucleotide sequence as shown in SEQ ID NO: 1 or a complementary sequence thereof, wherein position 51 of the sequence is a SNP site Chr1F: 20752, and the allele of the SNP site is C / A; Alternatively, the nucleic acid molecule comprises the nucleotide sequence shown in SEQ ID NO: 2 or its complementary sequence, wherein position 51 of the sequence is the SNP site Chr2B: 20889, and the allele of the SNP site is A / C; Alternatively, the nucleic acid molecule comprises the nucleotide sequence shown in SEQ ID NO: 3 or its complementary sequence, wherein position 51 of the sequence is the SNP site Chr3D: 384, and the allele of the SNP site is A / G; Alternatively, the nucleic acid molecule comprises the nucleotide sequence shown in SEQ ID NO: 4 or its complementary sequence, wherein position 51 of the sequence is a SNP site Chr4J: 17220, and the allele of the SNP site is G / T; Alternatively, the nucleic acid molecule comprises the nucleotide sequence shown in SEQ ID NO: 5 or its complementary sequence, wherein position 51 of the sequence is a SNP site Chr5D: 15964, and the allele of the SNP site is A / T; Alternatively, the nucleic acid molecule comprises the nucleotide sequence shown in SEQ ID NO: 6 or its complementary sequence, wherein position 51 of the sequence is the SNP site Chr6K: 15569, and the allele of the SNP site is A / C; Alternatively, the nucleic acid molecule comprises the nucleotide sequence shown in SEQ ID NO: 7 or its complementary sequence, wherein position 51 of the sequence is a SNP site Chr7D: 3376, and the allele of the SNP site is T / C; Alternatively, the nucleic acid molecule comprises the nucleotide sequence shown in SEQ ID NO: 8 or its complementary sequence, wherein position 51 of the sequence is the SNP site Chr8A: 19687, and the allele of the SNP site is G / A; Alternatively, the nucleic acid molecule comprises the nucleotide sequence shown in SEQ ID NO: 9 or its complementary sequence, wherein position 51 of the sequence is a SNP site Chr9H: 16936, and the allele of the SNP site is G / A; Alternatively, the nucleic acid molecule comprises a nucleotide sequence as shown in SEQ ID NO: 10 or its complementary sequence, wherein the 51st position of the sequence is a SNP site Chr10L: 4675, and the allele of the SNP site is T / C.

8. A primer or probe for detecting insect resistance in sugarcane, characterized in that: The target of the primer or probe is the isolated nucleic acid molecule of claim 7.

9. A kit for detecting insect resistance of sugarcane, characterized in that: The kit comprises: a primer or a probe for detecting the SNP site according to claim 1, or a primer or a probe for detecting insect resistance of sugarcane according to claim 8.

10. A SNP typing chip, characterized in that: The SNP site-specific probe according to claim 1 is fixed on the solid phase carrier of the chip.

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