A dCAPS marker related to wheat root length and drought resistance and its application

Abstract

This invention discloses a dCAPS marker related to wheat root length and drought resistance and its applications. This invention belongs to the field of biotechnology. The substance developed in this invention for detecting polymorphisms or genotypes at the InDel site in the wheat genome is applied to the following: (1) identifying or assisting in the identification of wheat root length and drought resistance; (2) screening or breeding wheat plants, lines, strains, or varieties with long root lengths and strong drought resistance; (3) screening or breeding wheat plants, lines, strains, or varieties with short root lengths and weak drought resistance; (4) wheat breeding; (5) preparing products for screening or breeding wheat plants, lines, strains, or varieties with long root lengths and drought resistance, wherein the InDel site is a site on wheat chromosome 3B, specifically nucleotides 19-21 of sequence 1 in the sequence listing. The dCAPS molecular marker of this invention can provide a strong selection signal, improving the accuracy of identifying wheat lateral root length and drought resistance.

Description

Technical Field

[0001] This invention belongs to the field of biotechnology, specifically relating to a dCAPS marker related to wheat root length and drought resistance and its application. Background Technology

[0002] Roots are the primary organs for wheat to absorb soil moisture and nutrients. A well-developed root system significantly enhances wheat's ability to absorb soil moisture and nutrients, effectively improving soil resource utilization. Drought stress promotes the extension of wheat roots into deeper soil layers. Under nutrient deficiency stress, wheat can increase root length to expand the contact area between the root system and the soil, thereby enhancing its ability to capture and absorb nutrients. However, since roots are hidden in the soil, their phenotypes are difficult to observe directly. The rapid development of molecular breeding technology has provided an opportunity for efficient root selection. Therefore, discovering and utilizing molecular markers related to root traits, and accelerating root architecture improvement through marker-assisted selection, is an urgent need for the sustainable development of wheat production. Summary of the Invention

[0003] The main problem this invention aims to solve is how to quickly identify wheat root length and drought resistance.

[0004] To address the aforementioned problems, this invention first provides the application of a substance for detecting polymorphisms or genotypes at the InDel site in the wheat genome in any of the following:

[0005] (1) To identify or assist in the identification of wheat root length and drought resistance; (2) Screening or breeding wheat single plants, lines, strains or varieties with long roots and strong drought resistance; (3) Screening or breeding wheat individual plants, strains, varieties or strains with short root length and weak drought resistance; (4) Wheat breeding; (5) Prepare products for identification or auxiliary identification of wheat root length and drought resistance; (6) Prepare or select wheat single plants, strains, varieties or cultivars with long roots and strong drought resistance; (7) Prepare or select wheat single plants, strains, varieties or cultivars with short root length and weak drought resistance; (8) Prepare products for wheat breeding; Using the genome sequence of wheat Chinese spring as a reference genome (https: / / plants.ensembl.org / Triticum_aestivum / Info / Index), the InDel site is a site on wheat chromosome 3B, specifically located at 627740215-627740217 bp, which is nucleotides 19-21 of sequence 1 in the sequence listing.

[0006] The present invention also provides a method for identifying or assisting in the identification of wheat root length and drought resistance, including detecting the genotype of the InDel site in the genome of the wheat to be tested, and identifying or assisting in the identification of wheat root length and drought resistance traits based on the genotype, wherein the InDel site is a site on wheat chromosome 3B, and is nucleotides 19-21 of sequence 1 in the sequence listing.

[0007] Furthermore, the genotype of InDel is In-AGT or Del-AGT, wherein the In-AGT genotype is a homozygous form of InDel containing nucleotides 19-21 of sequence 1, and the Del-AGT genotype is a homozygous form of InDel not containing nucleotides 19-21 of sequence 1. The identification or auxiliary identification of wheat root length and drought resistance based on the InDel genotype can be performed using any of the following methods: 1) The wheat to be tested with the InDel genotype In-AGT is or is a candidate wheat with long root length and strong drought resistance. 2) The wheat to be tested with the InDel genotype Del-AGT is or is a candidate wheat with short root length and weak drought resistance. 3) The root length of the wheat tested with the InDel genotype In-AGT is longer than that of the wheat tested with the InDel genotype Del-AGT, and its drought resistance is stronger than that of the wheat tested with the InDel genotype Del-AGT.

[0008] As one implementation scheme, the method for identifying or assisting in the identification of wheat root length and drought resistance includes the following steps: (1) Using the genomic DNA of the wheat to be tested as a template, PCR amplification was performed using the following primer combination; The primer composition consists of primers TaLAC69-dCAPS-F and TaLAC69-dCAPS-R; The primer TaLAC69-dCAPS-F is a single-stranded DNA molecule whose nucleotide sequence is sequence 2 in the sequence listing; The primer TaLAC69-dCAPS-R is a single-stranded DNA molecule whose nucleotide sequence is sequence 3 in the sequence listing.

[0009] (2) After completing step (1), the PCR product is treated with restriction endonuclease. Mbo II. Perform enzyme digestion, and then perform gel electrophoresis on the digestion products to determine the genotype of InDel in the wheat to be tested; (3) Identify the root length and drought resistance of the wheat to be tested based on the genotype results: The wheat samples with the InDel genotype In-AGT (with two fragments of 292 bp and 27 bp in size) exhibited longer root length and stronger drought resistance. The InDel genotype is Del-AGT (containing only a 316 bp fragment, i.e., not...). Mbo The wheat roots tested by enzyme digestion (II) showed short root length and weak drought resistance.

[0010] This invention also provides the application of the methods described above in wheat breeding.

[0011] This invention also provides a method for wheat breeding, wherein the method is M1 or M2. M1. The method includes detecting the genotype of InDel mentioned above in the wheat genome, selecting wheat with the genotype In-AGT of InDel as a parent for breeding, wherein In-AGT is a homozygous form of InDel containing nucleotides 19-21 of sequence 1, and the breeding purpose of the method includes selecting wheat with long roots and strong drought resistance. M2. The method includes detecting the genotype of InDel mentioned above in the wheat genome, selecting wheat with the genotype Del-AGT of InDel as a parent for breeding, wherein Del-AGT is the homozygous type of InDel that does not contain nucleotides 19-21 of sequence 1, and the breeding purpose of the method includes selecting wheat with short root length and weak drought resistance.

[0012] This invention also provides a product containing a substance for detecting polymorphisms or genotypes at the InDel site in the wheat genome, said product being any of the following: C1) Products that detect single nucleotide polymorphisms or genotypes related to wheat root length and drought resistance; C2) Products used to identify or assist in the identification of wheat root length and drought resistance; C3) Products used in wheat breeding; C4) Screening or breeding products of wheat single plants, strains, varieties or cultivars with long roots and strong drought resistance. C5) Screening or breeding of wheat single plants, strains, varieties or cultivars with short root lengths and weak drought resistance.

[0013] In the above applications, methods, and products, the substance may be a reagent and / or instrument required to determine the polymorphism or genotype of the SNP site by at least one of the following methods: DNA sequencing, restriction enzyme fragment color polymorphism, single-strand conformation polymorphism, denaturing high-performance liquid chromatography, and SNP chips. Among these, SNP chips include chips based on nucleic acid hybridization reactions, chips based on single-base extension reactions, chips based on allele-specific primer extension reactions, chips based on one-step reactions, chips based on primer ligation reactions, chips based on restriction endonuclease reactions, chips based on protein-DNA binding reactions, and chips based on fluorescent molecule-DNA binding reactions.

[0014] In this article, the substance may be D1), D2), or D3). D1) Contains a primer composition for amplifying wheat genomic DNA fragments including the InDel site; D2) PCR reagents containing the primer composition described in D1); D3) A kit containing the primer composition described in D1) or the PCR reagent described in D2).

[0015] In the above applications, methods, and products, the primer composition may or may not be labeled with a marker. The marker refers to any atom or molecule that can provide a detectable effect and can be linked to nucleic acids. Markers include, but are not limited to, dyes; radioactive markers, such as... 32 P; binding moieties, such as biotin; haptens, such as digoxigenin (DIG); luminescent, phosphorescent, or fluorescent moieties; and fluorescent dyes alone or in combination with moieties whose emission spectra can be inhibited or shifted by fluorescence resonance energy transfer (FRET). The label can provide a signal detectable by fluorescence, radioactivity, colorimetry, gravimetric determination, X-ray diffraction or absorption, magnetism, enzyme activity, etc. The label can be a charged moieties (positive or negative) or, optionally, charge-neutral. The label can include nucleic acid or protein sequences or combinations thereof, provided that the sequence containing the label is detectable. In some embodiments, nucleic acids are detected directly without labeling (e.g., direct sequence reading).

[0016] Furthermore, the primer composition consists of primer TaLAC69-dCAPS-F and primer TaLAC69-dCAPS-R; The primer TaLAC69-dCAPS-F is a single-stranded DNA molecule whose nucleotide sequence is sequence 2 in the sequence listing; The primer TaLAC69-dCAPS-R is a single-stranded DNA molecule whose nucleotide sequence is sequence 3 in the sequence listing.

[0017] The present invention also provides DNA molecules with nucleotide sequences as shown in Sequence 1 or Sequence 4 of the sequence listing.

[0018] This invention also provides the application of the above-mentioned InDel molecular markers in any of the following: (1) To identify or assist in the identification of wheat root length and drought resistance; (2) Screening or breeding wheat single plants, lines, strains or varieties with long roots and strong drought resistance; (3) Screening or breeding wheat individual plants, strains, varieties or strains with short root length and weak drought resistance; (4) Wheat breeding; (5) Prepare products for identification or auxiliary identification of wheat root length and drought resistance; (6) Prepare or select wheat single plants, strains, varieties or cultivars with long roots and strong drought resistance; (7) Prepare or select wheat single plants, strains, varieties or cultivars with short root length and weak drought resistance; (8) Prepare products for wheat breeding.

[0019] Optionally, in the above applications, the InDel site serves as a detection target.

[0020] The substance that detects the InDel site polymorphism and genotype can be combined with other substances (such as substances that detect single nucleotide polymorphisms or genotypes of other molecular markers related to wheat root length and drought resistance) to prepare a product for identifying wheat root length and drought resistance.

[0021] Experiments of this invention demonstrate that the dCAPS marker (PCR primer pair amplifying wheat genomic DNA fragments containing the dCAPS molecular marker), developed based on InDel (insertion / deletion) variants, can accurately genotype allelic variants related to root length and drought resistance, and predict wheat root length and drought resistance. Using marker-assisted selection breeding technology can effectively avoid the influence of environmental factors and human errors on phenotypic identification. The dCAPS molecular marker of this invention can provide a strong selection signal, improve the accuracy of identifying wheat lateral root length and drought resistance, and achieve the goal of marker-assisted selection for root length and drought resistance. Attached Figure Description

[0022] Figure 1 Agarose gel electrophoresis image of dCAPS-labeled wheat root length and drought resistance.

[0023] Figure 2 The total root length of different genotypes in 277 wheat materials was calculated.

[0024] Figure 3Association analysis of different genotypes and drought resistance in 277 wheat populations. , P <0.01.

[0025] Figure 4 The total root length of different genotypes in 99 wheat materials. Detailed Implementation

[0026] 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.

[0027] 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 and reagents used in the following examples are commercially available.

[0028] Unless otherwise specified, the quantitative experiments in the following examples are all repeated three times, and the results are averaged.

[0029] The 99 wheat-derived lines in the following examples are described in: Bai Yanming. Genetic analysis of diversity and bud-stage root drought resistance traits of wheat, white wheat and their derivatives. Taigu: Shanxi Agricultural University, 2019. This biological material is available to the public from the applicant and is intended solely for the replication of experiments of this invention and may not be used for any other purpose.

[0030] The 277 materials (12 local varieties, 230 modern bred varieties, and 35 high-generation lines) in the following examples have been described in: Li L, Mao X, Wang J, Chang X, Reynolds M, Jing R. Genetic dissection of drought and heat-responsive agronomic traits in wheat. Plant Cell and Environment, 2019, 42(9): 2540-2553. This biological material is available to the public from the applicant and is intended solely for the replication of experiments of this invention and may not be used for any other purpose.

[0031] The following examples used statistical software to process the data. The experimental results are expressed as mean ± standard deviation. One-way ANOVA was used, and P < 0.05 was considered satisfactory. () indicates a significant difference, P < 0.01. () indicates a highly significant difference.

[0032] Example 1: Localization of InDel sites related to wheat root length and drought resistance, and establishment of dCAPS molecular marker and detection method. The sample to be tested consisted of 277 materials (12 local varieties, 230 modern bred varieties and 35 high-generation lines) mainly from the late-maturing winter wheat region in northern my country and the Huang-Huai winter wheat region.

[0033] 1. Identification of wheat seedling root morphology: Thirty-five plump, uniform seeds were selected from each sample, treated with 10% sodium hypochlorite solution for 10 min, washed five times with distilled water, and cultured in petri dishes at 25℃ in the dark for 24 h. Seeds with good germination and uniform growth were transferred to culture boxes, and modified Hoagland nutrient solution (NSP1020, Beijing Coolplay Technology Co., Ltd.) was added. The plants were then cultured in an artificial climate chamber with a day / night temperature of 22℃ / 18℃, a photoperiod of 16 h / 8 h, and a light intensity of 150 μmol·m⁻¹. -2 ·s -1 Phenotypic results were observed on day 7. The root system was scanned using an Epson 10000XL scanner (Epson, Japan), and the root images were analyzed using WinRHIZO (2009a, Regent Instruments Ins, Canada) software. The total root length was read, and the results are shown in Table 1.

[0034] 2. Study on drought resistance phenotypes in wheat At the Shunyi Experimental Base of the Institute of Crop Science, Chinese Academy of Agricultural Sciences (40°23′N, 116°56′E), two treatments were established: dryland and irrigated land. The dryland was rainfed, while the irrigated land was irrigated (750 m²) before overwintering, during the heading and flowering stages. 3 hm -2 Each variety was sown in 4 rows, with a row length of 2 m, a row spacing of 30 cm, and 40 seeds per row. Conventional field management was applied. Plot yields were measured. The drought resistance coefficient was calculated as the ratio of the measured value of a specific trait under drought stress to the corresponding trait measured under normal irrigation (control) conditions. The formula was: Drought resistance coefficient = Trait value under drought stress / Trait value under normal irrigation. The results are shown in Table 1.

[0035] 3. Development of dCAPS molecular markers 1) Extract genomic DNA from the wheat to be tested; 2) PCR amplification Using wheat genomic DNA as a template, PCR amplification was performed on the fragment near the InDel molecular marker using the upstream primer TaLAC69-dCAPS-F: 5'-TGATTGCTGCTCAGGAAG -3' (sequence 2) and the downstream primer TaLAC69-dCAPS-R: 5'-CTACATGTTGGCATACATATTT -3' (sequence 3).

[0036] The PCR amplification system consisted of: 5 μL Taq DNA Polymerase and 0.4 μL primer F / R (10 μmol·L⁻¹). -1 1 μL of DNA template (200 ng·μL) -1 ) and ddH2O 3.2 μL; The PCR amplification program was as follows: 95℃ for 3 min; 95℃ for 15 s, 60℃ for 15 s, 72℃ for 30 s, 40 cycles; 72℃ for 3 min.

[0037] 3) PCR products were detected using 1.2% agarose gel electrophoresis, and the target bands were recovered, purified, and sequenced. The sequencing results were compared using DNAMAN software to analyze sequence polymorphism.

[0038] Based on sequencing results analysis, an InDel locus associated with wheat root length and drought resistance was identified. The molecular marker is located at 627740215-627740217 bp of wheat reference genome 3B, specifically at positions 19-21 of sequence 1 (base "AGT"). Wheat with the InDel locus type In-AGT exhibits long root length and drought resistance, and the wheat genotype with the InDel locus type In-AGT is defined as In-AGT; wheat with the InDel locus type Del-AGT exhibits short root length and is not drought resistant, and the wheat genotype with the InDel locus type Del-AGT is defined as Del-AGT. The genotyping results of 277 wheat accessions are shown in Table 1.

[0039] 4. The population was scanned using the developed dCAPS molecular marker.

[0040] 1) Extract genomic DNA from the wheat to be tested; 2) PCR amplification Using wheat genomic DNA as a template, PCR amplification was performed on the fragment near the InDel site using the upstream primer TaLAC69-dCAPS-F: 5'-TGATTGCTGCTCAGGAAG -3' (sequence 2) and the downstream primer TaLAC69-dCAPS-R: 5'-CTACATGTTGGCATACATATTT -3' (sequence 3).

[0041] The PCR amplification system consisted of: 5 μL Taq DNA Polymerase and 0.4 μL primer F / R (10 μmol·L⁻¹). -1 1 μL of DNA template (200 ng·μL) -1 ) and ddH2O 3.2 μL; The PCR amplification program was as follows: 95℃ for 3 min; 95℃ for 15 s, 60℃ for 15 s, 72℃ for 30 s, 40 cycles; 72℃ for 3 min.

[0042] 3) Enzyme digestion PCR amplification products were digested with restriction endonuclease MboII, and the digested products were detected by agarose gel electrophoresis. The digestion system was: MboII 0.3 μL, PCR product 2 μL, buffer 1 μL, ddH2O 6.7 μL; incubated at 37℃ for 2 h.

[0043] The experimental results (Table 1) show that, after amplification using primers TaLAC69-dCAPS-F and TaLAC69-dCAPS-R, the fragment lengths of the amplified products exhibited polymorphism after enzyme digestion. Two banding patterns were observed in 277 samples. Figure 1 Band type 1 consists of a band above 300 bp; band type 2 consists of a band below 300 bp. Sequencing revealed that the band above 300 bp in band type 1 is 316 bp (sequence 4); the band below 300 bp in band type 2 is 292 bp (positions 28-319 of sequence 1).

[0044] Table 1. Results of dCAPS molecular markers and root length drought resistance phenotypes in 277 materials.

[0045] The relationship between wheat root length and drought resistance index and dCAPS molecular marker banding was analyzed (Table 2). The results showed that: 1) The dCAPS molecular marker banding pattern of wheat with long roots (average root length of 53.71) is band type II, containing two bands of 292 bp and 27 bp (i.e., the genotype of the InDel site is In-AGT); the dCAPS molecular marker banding pattern of wheat with short roots (average root length of 47.84) is band type I, i.e., the enzyme digestion product is only one band of 316 bp (i.e., the genotype of the InDel site is Del-AGT).

[0046] 2) Wheat dCAPS molecular marker banding pattern with strong drought resistance (average yield drought resistance coefficient of 0.86) is band type II containing two bands of 292 bp and 27 bp (i.e., the genotype of the InDel site is In-AGT); wheat dCAPS molecular marker banding pattern with weak drought resistance (average yield drought resistance coefficient of 0.83) is band type I, i.e., the enzyme digestion product is only one band of 316 bp (i.e., the genotype of the InDel site is Del-AGT).

[0047] Table 2. Statistical results of dCAPS molecular markers and root length drought resistance phenotypes in 277 materials.

[0048] Based on the root system analysis results of 277 wheat varieties, the average total root length and average yield drought resistance coefficient of wheat germplasm with an electrophoretic band size of 292 bp (genotype In-AGT) were greater than those of wheat germplasm with an electrophoretic band size of 316 bp (genotype Del-AGT). Figure 2 and Figure 3 ).

[0049] Example 2: Application of dCAPS markers in assisted selection of long-rooted wheat germplasm Samples to be tested: 99 wheat derivative lines.

[0050] 1. Method for determining the total root length of wheat seedlings The method was the same as in Example 1. The total root length of 99 wheat-derived line populations was identified, and the results are shown in Table 3 below.

[0051] 2. dCAPS marker detection Total genomic DNA was extracted from leaves of 99 wheat materials at the three-leaf stage. Using the total genomic DNA as a template, PCR amplification was performed using forward primer TaLAC69-dCAPS-F and reverse primer TaLAC69-dCAPS-R. The PCR amplification and enzyme digestion reaction system and conditions were the same as in Example 1. The results are shown in Table 3.

[0052] Table 3. dCAPS marker detection results and total root length results

[0053] The total root length of 99 wheat white wheat derivatives was measured, and the results are as follows: Figure 4 As shown: Analysis of the total root length of 99 wheat-derived lines showed that the average total root length of the materials with an amplified product sequence length of 316 bp (band type 1) was 50.77 cm, while the average total root length of the materials with amplified product sequence lengths of 297 bp and 27 bp (band type 2) was 59.1 cm. The average total root length of wheat materials containing band type 2 (electrophoretic band size of 292 bp) was greater than that of wheat materials containing band type 1 (electrophoretic band size of 316 bp).

[0054] In summary, the dCAPS molecular marker and specific primer sequences developed in this invention can be directly used in marker-assisted selection breeding to identify or assist in the identification of wheat root length and drought resistance.

[0055] The present invention has been described in detail above. Those skilled in the art will recognize that 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. While specific embodiments have been provided, 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.

Claims

1. The application of substances used to detect polymorphisms or genotypes at the InDel site in the wheat genome in any of the following: (1) To identify or assist in the identification of wheat root length and drought resistance; (2) Screening or breeding wheat single plants, lines, strains or varieties with long roots and strong drought resistance; (3) Screening or breeding wheat individual plants, strains, varieties or strains with short root length and weak drought resistance; (4) Wheat breeding; (5) Prepare products for identification or auxiliary identification of wheat root length and drought resistance; (6) Prepare or select wheat single plants, strains, varieties or cultivars with long roots and strong drought resistance; (7) Prepare or select wheat single plants, strains, varieties or cultivars with short root length and weak drought resistance; (8) Prepare products for wheat breeding; The InDel site is a site on wheat chromosome 3B, specifically nucleotides 19-21 of sequence 1 in the sequence listing.

2. A method for identifying or assisting in the identification of wheat root length and drought resistance, characterized in that: This includes detecting the genotype of the InDel site in the wheat genome, and identifying or assisting in the identification of wheat root length and drought resistance traits based on the genotype. The InDel site is a site on wheat chromosome 3B, and the site on chromosome 3B is nucleotides 19-21 of sequence 1 in the sequence listing.

3. The application according to claim 1 or the method according to claim 2, characterized in that: The InDel genotype is either In-AGT or Del-AGT, where the In-AGT genotype is a homozygous form of InDel containing nucleotides 19-21 of sequence 1, and the Del-AGT genotype is a homozygous form of InDel not containing nucleotides 19-21 of sequence 1. The identification or auxiliary identification of wheat root length and drought resistance based on the InDel genotype can be performed using any of the following methods: 1) The wheat to be tested with the InDel genotype In-AGT is or is a candidate wheat with long root length and strong drought resistance. 2) The wheat to be tested with the InDel genotype Del-AGT is or is a candidate wheat with short root length and weak drought resistance. 3) The root length of the wheat tested with the InDel genotype In-AGT is longer than that of the wheat tested with the InDel genotype Del-AGT, and its drought resistance is stronger than that of the wheat tested with the InDel genotype Del-AGT.

4. The application of the method according to claim 2 or 3 in wheat breeding.

5. A method for wheat breeding, characterized by: The method is M1 or M2. M1. The method includes detecting the genotype of InDel in claim 1 in the wheat genome, selecting wheat with the genotype In-AGT of InDel as a parent for breeding, wherein In-AGT is a homozygous form of InDel containing nucleotides 19-21 of sequence 1, and the breeding purpose of the method includes selecting wheat with long roots and strong drought resistance. M2. The method includes detecting the genotype of InDel as described in claim 1 in the wheat genome, selecting wheat with the genotype Del-AGT of InDel as a parent for breeding, wherein Del-AGT is a homozygous form of InDel that does not contain nucleotides 19-21 of sequence 1, and the breeding purpose of the method includes selecting wheat with short root length and weak drought resistance.

6. A product containing a substance for detecting polymorphisms or genotypes at the InDel site in the wheat genome, characterized in that: The product is any one of the following: C1) Products that detect single nucleotide polymorphisms or genotypes related to wheat root length and drought resistance; C2) Products used to identify or assist in the identification of wheat root length and drought resistance; C3) Products used in wheat breeding; C4) Screening or breeding products of wheat single plants, strains, varieties or cultivars with long roots and strong drought resistance. C5) Screening or breeding of wheat single plants, strains, varieties or cultivars with short root lengths and weak drought resistance.

7. The application according to claim 1 or the product according to claim 6, characterized in that: The substance is either D1), D2), or D3). D1) Contains a primer composition for amplifying wheat genomic DNA fragments including the InDel site; D2) PCR reagents containing the primer composition described in D1); D3) A kit containing the primer composition described in D1) or the PCR reagent described in D2).

8. The application, method, or product according to claim 7, characterized in that: The primer composition consists of primers TaLAC69-dCAPS-F and TaLAC69-dCAPS-R; The primer TaLAC69-dCAPS-F is a single-stranded DNA molecule whose nucleotide sequence is sequence 2 in the sequence listing; The primer TaLAC69-dCAPS-R is a single-stranded DNA molecule whose nucleotide sequence is sequence 3 in the sequence listing.

9. A DNA molecule, characterized by: The nucleotide sequences are shown in Sequence 1 or Sequence 4 of the sequence listing.

10. The use of the InDel molecular marker of claim 1 in any of the following: (1) To identify or assist in the identification of wheat root length and drought resistance; (2) Screening or breeding wheat single plants, lines, strains or varieties with long roots and strong drought resistance; (3) Screening or breeding wheat individual plants, strains, varieties or strains with short root length and weak drought resistance; (4) Wheat breeding; (5) Prepare products for identification or auxiliary identification of wheat root length and drought resistance; (6) Prepare or select wheat single plants, strains, varieties or cultivars with long roots and strong drought resistance; (7) Prepare or select wheat single plants, strains, varieties or cultivars with short root length and weak drought resistance; (8) Prepare products for wheat breeding.

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