CN115820897B - Molecular marker closely linked with corn female spike and sword leaf length and application thereof - Google Patents
Molecular marker closely linked with corn female spike and sword leaf length and application thereof Download PDFInfo
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Abstract
The invention discloses a molecular marker closely linked with the length of corn female spike sword leaf and application thereof. The molecular marker closely linked with the length of the corn female spike sword leaf is a DNA fragment shown as SEQ ID No.3 or a DNA fragment shown as SEQ ID No.4, and can be detected by using a primer pair consisting of two single-stranded DNAs shown as SEQ ID No.1 and SEQ ID No.2 in a sequence table. Experiments prove that the molecular marker closely linked with the corn female ear sword leaf length is related to the corn female ear sword leaf length, the female ear sword leaf length condition of corn can be successfully identified by utilizing the molecular marker, and the molecular marker has the advantages of simplicity, convenience, rapidness, high efficiency and accuracy, and high repeatability and specificity, can be used for corn molecular marker assisted breeding, and can be used for breeding new corn varieties with excellent comprehensive properties, so that the breeding cost is greatly saved, and the breeding efficiency is improved.
Description
Technical Field
The invention relates to a molecular marker closely linked with the length of female corn ear sword leaf and application thereof in the field of biotechnology.
Background
Corn (Zea mays l.) is a dual-purpose crop for food, feed and economy, and is the only crop in our country that steadily increases in sowing area and yield. In recent years, global economy has rapidly developed, and demand for corn is expected to increase in future industrial production and life. The planting area of the corn in 2021 is 4332 ten thousand hectares, which is increased by 206 ten thousand hectares compared with the last year, the yield is up to 27255 ten thousand tons, and the yield is about 39.91 percent of the total yield of the grain in China. Therefore, corn production has very important strategic position in guaranteeing the grain safety of China.
The sword leaf is a further extension of the top end of the maize female ear bud, is an organ for degeneration in the long-term evolution process of maize, and is one of important characters for identifying maize varieties and germplasm sources. The length of the swords of corn of different germplasm sources varies greatly, most temperate sweet corn has longer swords, while most tropical corn has no or shorter swords. The sword leaf is used as an important component of the corn female ear, and has important effects on the growth and development of the corn female ear and the formation of the yield. Therefore, the genetic basis of the length of the female spike sword leaf of the corn is further analyzed, and the molecular marker closely linked with the number character locus (quantitative trait loci, QTL) of the length of the female spike sword leaf is developed, so that the method has important significance for cultivating new excellent corn varieties which are suitable in the length character of the female spike sword leaf and have larger market economic value.
The molecular markers have the advantages of large quantity, no influence of environmental conditions, development period, expression regulation and control and other factors, capability of providing complete and rich genetic information and the like, and are widely applied to aspects of germplasm resource identification, QTL positioning, molecular marker auxiliary selection and the like. InDel (InDel) markers are one of the commonly used molecular markers based on differences in DNA levels, specifically differences in two materials, a certain number of nucleotide insertions or deletions in certain loci of the genome of one material relative to the other, and PCR primers are designed to amplify these InDel sites based on their InDel sites. By utilizing InDel markers closely linked with target genes, assisted backcross selection, assisted pedigree selection and even whole genome selection are adopted, so that linkage encumbrance is reduced, favorable genes are polymerized, the breeding process is accelerated, and the selection efficiency and effect can be effectively improved.
At present, although research reports on QTL for controlling the length of corn female spike and sword leaf exist, no molecular marker closely linked with a target QTL is developed and applied for patent application, and no patent report related to the length of corn female spike and sword leaf exists in the qFL3-1 segment.
Disclosure of Invention
The invention aims to solve the technical problem of how to detect the length characters of the female spike and sword leaf of corn.
In order to solve the technical problems, the invention firstly provides application of a substance for detecting the corn female spike and sword leaf length molecular marker in detecting or assisting in detecting the corn female spike and sword leaf length character, wherein the corn female spike and sword leaf length molecular marker is a DNA fragment shown as SEQ ID No.3 and a DNA fragment shown as SEQ ID No. 4.
In the application, the substance for detecting the molecular marker of the length of the female ear and sword leaf of the corn can be a primer pair consisting of two single-stranded DNA shown as SEQ ID No.1 and SEQ ID No.2 in a sequence table.
The invention also provides a method for detecting the length characters of the female corn ear sword leaf, which comprises the following steps: the genome DNA of corn to be detected is used as a template, a primer pair consisting of two single-stranded DNA shown as SEQ ID No.1 and SEQ ID No.2 in a sequence table is utilized for PCR amplification, the length of the female spike sword leaf of the homozygous corn with the obtained PCR product sequence of SEQ ID No.3 is shorter or the length of the female spike sword leaf of the homozygous corn with the obtained PCR product sequence of SEQ ID No.4 is shorter or the length of the female spike sword leaf of the homozygous corn with the obtained PCR product sequence of SEQ ID No.3 is shorter or the length of the female spike sword leaf of the heterozygous corn with the obtained PCR product sequences of SEQ ID No.3 and SEQ ID No.4 is shorter or the length of the female spike sword leaf of the heterozygous corn with the obtained PCR product sequence of SEQ ID No.4 is shorter or the candidate.
The invention also provides a method for detecting the length characters of the female corn ear sword leaf, which comprises the following steps: the genome DNA of the corn to be detected is used as a template, a primer pair consisting of two single-stranded DNA shown as SEQ ID No.1 and SEQ ID No.2 in a sequence table is used for carrying out PCR amplification, the length of the female spike sword leaf of the homozygous corn with the obtained PCR product size of 178bp is shorter than or candidate shorter than the length of the female spike sword leaf of the homozygous corn with the obtained PCR product size of 363bp, the length of the female spike sword leaf of the homozygous corn with the obtained PCR product size of 178bp is shorter than or candidate shorter than the length of the female spike sword leaf of the heterozygous corn with the obtained PCR product sizes of 178bp and 363bp, and the length of the female spike sword leaf of the heterozygous corn with the obtained PCR product size of 178bp and 363bp is shorter than or candidate shorter than the length of the female spike sword leaf of the homozygous corn with the obtained PCR product size of 363 bp.
The invention also provides a corn breeding method, which comprises the following steps: and (3) taking genome DNA of the corn to be detected as a template, carrying out PCR amplification by using a primer pair consisting of two single-stranded DNA shown as SEQ ID No.1 and SEQ ID No.2 in a sequence table, and selecting the corn to be detected with a PCR product of SEQ ID No.3 as a parent to finish breeding.
In the above, the reaction system for PCR amplification using the primer pair consisting of two single-stranded DNAs shown in SEQ ID No.1 and SEQ ID No.2 of the sequence Listing may be: 1. Mu.L of single-stranded DNA shown in SEQ ID No.1 at a concentration of 10. Mu. Mo 1/L; 1. Mu.L of single-stranded DNA shown in SEQ ID No.2 at a concentration of 10. Mu. Mo 1/L; 1. Mu.L of genomic DNA at a concentration of 100 ng/. Mu.L; 5 μL 2× Taq PCR StarMix with Loading Dye (Beijing Kang Runcheng Biotechnology Co., ltd., product number: A012-01); 2 μL ddH 2 O.
The reaction conditions for PCR amplification using a primer pair consisting of two single-stranded DNAs shown as SEQ ID No.1 and SEQ ID No.2 in the sequence Listing may be: pre-denaturation at 95℃for 10min; denaturation at 95℃for 45s, annealing at 57℃for 45s, elongation at 72℃for 35s,35 cycles; extending at 72 ℃ for 10min; preserving at 4 ℃.
The application of the substance for detecting the molecular marker of the corn female spike and sword leaf length in preparing a product for detecting the corn female spike and sword leaf length property also belongs to the protection scope of the invention.
The application of the corn female spike and sword leaf length molecular marker in detecting or assisting in detecting the corn female spike and sword leaf length character also belongs to the protection scope of the invention.
The application of the corn female ear and sword leaf length molecular marker in corn breeding also belongs to the protection scope of the invention.
In the invention, the length of the female spike sword leaf can be the length which further extends outwards from the top end of the corn female spike bud leaf.
The corn female spike and sword leaf length molecular marker is related to the corn female spike and sword leaf length, can be used for successfully identifying the corn female spike and sword leaf length condition, has the advantages of simplicity, convenience, rapidness, high efficiency and accuracy, is good in repeatability and high in specificity, can be used for corn molecular marker assisted breeding, and can be used for breeding new corn varieties with excellent comprehensive properties, the breeding cost is greatly saved, and the breeding efficiency is improved.
The following detailed description of the invention is provided in connection with the accompanying drawings that are presented to illustrate the invention and not to limit the scope thereof. The examples provided below are intended as guidelines for further modifications by one of ordinary skill in the art and are not to be construed as limiting the invention in any way.
Drawings
FIG. 1 shows the result of the molecular marker qFL3-1 of the present invention in the alignment of amplified sequences in parents.
FIG. 2 is an electropherogram of the PCR amplified product of the molecular marker qFL3-1 of the present invention in parents. Wherein W is the amplified band type of the maize inbred line W22, C is the amplified band type of the maize wild inbred species CIMMYT 8759, and the Marker has the band sizes of 100bp, 250bp, 500bp, 750bp, 1000bp, 1500bp and 2000bp from bottom to top.
FIG. 3 is an electropherogram of the PCR amplified product of the molecular marker qFL3-1 of the present invention in the F 2 population. Wherein, W is the amplified band type of homozygous W22 genotype, C is the amplified band type of homozygous CIMMYT 8759 genotype, H is the amplified band type of heterozygous genotype, and the Marker sizes are 100bp, 250bp, 500bp, 750bp, 1000bp, 1500bp and 2000bp from bottom to top.
FIG. 4 is a single marker analysis result of the length trait of female spike sword leaf in the F 2 population of the molecular marker qFL3-1 of the present invention. Nil_w22 represents homozygous W22 genotype, het represents heterozygous genotype, nil_cimyt 8759 represents homozygous cimyt 8759 genotype, x represents very significant difference (P < 0.01).
Detailed Description
The experimental methods in the following examples, unless otherwise specified, are conventional methods, and are carried out according to techniques or conditions described in the literature in the field or according to the product specifications. Materials, reagents, instruments and the like used in the examples described below are commercially available unless otherwise specified. The quantitative tests in the following examples were all set up in triplicate and the results averaged. In the following examples, unless otherwise specified, the 1 st position of each nucleotide sequence in the sequence listing is the 5 'terminal nucleotide of the corresponding DNA/RNA, and the last position is the 3' terminal nucleotide of the corresponding DNA/RNA.
The maize inbred line W22 and the maize wild-type inbred species CIMMYT8759 in the examples described below are both described in document (Identification and fine mapping of quantitative trait loci for the number of vascular bundle in maize stem,JIntegrPlant Biol.2016Jan;58(1):81-90.doi:10.1111/jipb.12358.Epub 2015Jul 16.), the maize wild-type inbred species CIMMYT8759 being CIMMYT access 8759 in this document, the maize inbred line W22 and the maize wild-type inbred species CIMMYT8759 being available to the public from the applicant, which biological material was used only for repeated experiments related to the invention and not for other uses.
MR0242 in the following examples is a U.S. corn germplasm resources center (Maize Genetics Cooperation Stock Center) product with the website: www.maizecoop.cropsci.uiuc.edu the public may also obtain this biomaterial from the applicant, which was only used for repeated experiments in connection with the invention and not as a further use. MR0242 is an introgressed material derived from maize inbred line W22 by crossing, backcrossing and selfing with maize wild inbred CIMMYT 8759.
Example 1: molecular marker closely linked with corn female spike and sword leaf length
The invention provides a molecular marker qFL3-1 (marked as corn female spike and sword leaf length molecular marker) for identifying or assisting in identifying corn female spike and sword leaf length, wherein the molecular marker is a DNA fragment obtained by PCR (polymerase chain reaction) amplification of a primer pair A1 by taking corn genome DNA as a template, and the sequence of the obtained DNA fragment is SEQ ID No.3 or SEQ ID No.4. The primer set A1 has the following sequence:
forward amplification primer FL3-F:5'-GCCCCTGCGATGTTCAAC-3' as shown in SEQ ID No. 1;
reverse amplification primer FL3-R:5'-AGAATTCCAGTTGCTTAAGAGGC-3' as shown in SEQ ID No. 2;
The sequence of the obtained PCR product is detected by taking the genome DNA of a corn inbred line W22 with shorter female spike and sword leaf length and a corn wild kindred species CIMMYT8759 with longer female spike and sword leaf length as templates, and carrying out PCR amplification by using a forward amplification primer shown as SEQ ID No.1 and a reverse amplification primer shown as SEQ ID No. 2.
Wherein, the reaction system of 10 mu L PCR amplification is as follows:
(1) 1. Mu.L of forward amplification primer shown in SEQ ID No.1 at a concentration of 10. Mu.m 1/L;
(2) 1. Mu.L of the reverse amplification primer shown in SEQ ID No.2 at a concentration of 10. Mu.m 1/L;
(3) 1. Mu.L of DNA template at a concentration of 100 ng/. Mu.L;
(4) 5 μL 2× Taq PCR StarMix with Loading Dye (Beijing Kang Runcheng Biotechnology Co., ltd., product number: A012-01);
(5)2μL ddH2O。
The procedure for PCR amplification was as follows:
(1) Pre-denaturation at 95℃for 10min;
(2) Denaturation at 95℃for 45s, annealing at 57℃for 45s, elongation at 72℃for 35s,35 cycles;
(3) Extending at 72 ℃ for 10min;
(4) Preserving at 4 ℃.
PCR instrument model: eppendorf Mastercycler nexus.
The PCR amplified product is subjected to electrophoresis separation in 2.0% agarose gel (each 100mL of gel solution contains 2.0g agarose), sequencing analysis is carried out, and the result shows that the PCR amplified product is subjected to PCR amplification by taking genomic DNA of a maize inbred line W22 as a template, the molecular weight of the PCR amplified product is 178bp, and the nucleotide sequence of the PCR amplified product is shown as SEQ ID No. 3; the genome DNA of wild kindred species CIMMYT 8759 of corn is used as template to make PCR amplification, the molecular weight of PCR amplification product is 363bp, and its nucleotide sequence is shown in SEQ ID No. 4.
SEQ ID No.3：
GCCCCTGCGATGTTCAACATACTGCTCATAGGCATAACTTGAGGGTGTGTGGGGCTTATCATAGCACCATTGGATGGATCTCAAAATTTAAAATTTGGCGCACATATTGCATCATGGAATACTATATTACCGTGTTCAGCATCTATACTCTTTGTGCCTCTTAAGCAACTGGAATTCT.
SEQ ID No.4：
GCCCCTGCGATGTTCAACATACTGCTCATAGGCATAACTTGAGGGTGTGTGGGGCTTATCATAGCACCATTGGATGCAGGGGCGAAGCCAGCATTTAAGATTGAGAGGGACAAATTAGATTGAGGGGGGCTGTTAAGAGGTATTTTACATTATTTATATGGTGATTAGCTAAAAAAATTAGTAGCTTCTATGGAATTTGTAGAAGATTAGGGGGGACATTGCCCCCCTGTGCCCCTCCCTAGAATCGCCCCTGATTGGATGGATCTCAAAATTTAAAATTTGGCACACATATTGCATCATGGAATACTATATTACCGTGTTCAGCATCTATACTCTTTGTGCCTCTTAAGCAACTGGAATTCT.
Wherein, the amplified band of the maize inbred line W22 is an allele for shortening the length of the female spike sword leaf. Therefore, if the molecular weight of the PCR amplified product of the corn sample to be detected is 178bp, the corn sample to be detected contains alleles for shortening the length of the female ear and sword leaf of corn; if the molecular weight of the PCR amplified product of the corn sample to be detected is 363bp, the corn sample to be detected contains the allele for increasing the length of the female spike and sword leaf of corn.
The alignment results of SEQ ID No.3 and SEQ ID No.4 are shown in FIG. 1. The electrophoresis results of PCR amplified products of maize inbred line W22 and maize wild-type inbred CIMMYT 8759 are shown in FIG. 2.
Example 2: method for obtaining molecular marker FL3
The method for obtaining the molecular marker FL3 specifically comprises the following steps:
Step 1: construction of BC 2S3 introgression line populations containing 866 lines
Corn inbred line W22 is used as an acceptor parent, corn wild inbred species CIMMYT 8759 is used as a donor parent, and the BC 2S3 introgression line population containing 866 families is obtained through hybridization for 1 generation, backcross for 2 generation and 3 generation inbreeding.
Step 2: field planting and phenotyping of introgressed populations
BC 2S3 infiltration line populations were planted in the spring 2019 at the national crop variety area test station in Liuyang City (28.2N, 113.6E) of Hunan province. The field test adopts an amplified incomplete random granule design. 2 rows of plants are planted in each district, 15 plants are planted in each row, and the plant spacing is 25cm. 2 families are planted in each ridge. The ridge height is 15cm, the ridge width is 70cm, and the ditch width is 30cm.
After 10 days of powder dispersion, the female spike sword leaf length phenotype was measured, and 10 individual plants were continuously investigated for each family material starting from strain 2. The length of the female spike sword leaf is the length which further extends outwards from the top end of the corn female spike bract.
Step 3: performing QTL positioning analysis
And carrying out QTL positioning analysis by utilizing a multi-QTL model of R/QTL. Firstly, performing simple interval positioning analysis on the QTL by using Haley-Knott regression, and determining the LOD threshold value (alpha=0.05) of the female spike sword-leaf length QTL by adopting a method of substitution test 10000 times. And (3) carrying out multi-QTL model fitting on the QTL model obtained by positioning the simple interval, and optimizing the position of each QTL by utilizing a refineqtl command of R/QTL. Further utilizing addqtl command to detect if other significant improvement model QTL exists in genome, if new QTL is detected, re-fitting multi QTL model and optimizing QTL position, repeating this process until new QTL is not detected. Finally, the fitqtl command is used for calculating the total surface type variation of all QTL interpretations and the additive effect and the phenotype contribution rate of the single QTL.
QTL positioning result analysis: a total of 9 QTL's controlling the length of the maize female ear and leaf were detected, of which one QTL qFL3-1 with the greatest phenotypic effect was detected on chromosome 3. The LOD value of qFL3-1 is 25.31, the additive effect size is 1.14cm, the dominant effect size is 0.28cm, the phenotype contribution rate is 10.48%, and the phenotype contribution rate is located in a range from 132255901bp to 151202327bp of a 3 rd chromosome of corn.
Step 4: development and synthesis of molecular marker FL3
Searching qFL3-1 in a physical interval of 132255901bp to 151202327bp of a 3 rd chromosome by utilizing on-line primer design software primer3 (https:// primer3.Ut. Ee /), and designing a forward amplification primer FL3-F and a reverse amplification primer FL3-R, wherein the primers are synthesized by Beijing qing department biotechnology Co., ltd, and the nucleotide sequence is as follows:
forward amplification primer FL3-F:5'-GCCCCTGCGATGTTCAAC-3' as shown in SEQ ID No. 1;
reverse amplification primer FL3-R:5'-AGAATTCCAGTTGCTTAAGAGGC-3' as shown in SEQ ID No. 2.
Example 3: application of molecular marker FL3
Introgression line MR0242, which is heterozygous only at the qFL3-1 segment and homozygous at the other genomic locus, was used as starting material and self-pollinated to generate a population of F 2 isolated only at the qFL3-1 segment. The molecular marker FL3 obtained by the invention is verified by taking F 2 groups containing 584 individual plants as materials, so as to determine the accuracy of the molecular marker applied to molecular marker assisted selective breeding. The method specifically comprises the following steps:
Step 1: determination of F 2 group female spike Sword leaf Length
The female spike sword leaf length of the F 2 population plants was determined as in example 2.
Step 2: the DNA of corn leaves is extracted by adopting a CTAB method.
Step 3: PCR amplification
The reaction system for PCR amplification was 10. Mu.L, including:
(1) 1. Mu.L of forward amplification primer shown in SEQ ID No.1 at a concentration of 10. Mu.m 1/L;
(2) 1. Mu.L of the reverse amplification primer shown in SEQ ID No.2 at a concentration of 10. Mu.m 1/L;
(3) 1. Mu.L of DNA template at a concentration of 100 ng/. Mu.L;
(4) 5 μL 2× Taq PCR StarMix with Loading Dye (Beijing Kang Runcheng Biotechnology Co., ltd., product number: A012-01);
(5)2μL ddH2O。
The procedure for PCR amplification was as follows:
(1) Pre-denaturation at 95℃for 10min;
(2) Denaturation at 95℃for 45s, annealing at 57℃for 45s, elongation at 72℃for 35s,35 cycles;
(3) Extending at 72 ℃ for 10min;
(4) Preserving at 4 ℃.
PCR instrument model: eppendorf Mastercycler nexus.
Step 4: electrophoresis
The electropherogram of the PCR amplified product of the molecular marker FL3 in part F 2 of the individual strain is shown in FIG. 3.
Step 5: analysis of results
Determining the genotype of the corn sample to be detected according to the molecular weight of the PCR amplification product: if the PCR amplified product of the corn sample to be tested has only one 178bp band, the corn to be tested is homozygous for the W22 genotype (i.e. the genotype identical to the corn inbred line W22); if the PCR amplified product of the corn sample to be detected has only one 363bp band, the corn to be detected is homozygous CIMMYT 8759 genotype (namely the genotype same as the wild kindred species CIMMYT 8759 of corn); if the PCR amplified product of the corn sample to be detected has not only a 178bp band but also a 363bp band, the corn to be detected is heterozygous genotype.
The total 138 strains in the F 2 single strain are homozygous W22 genotypes, and sequencing of PCR products shows that the sequences of the F 2 single strain are SEQ ID No.3, and the length of female spike sword leaves of the F 2 single strain with the 138 homozygous W22 genotypes is 0.43+/-0.79 cm; the total 144 strains are homozygous CIMMYT8759 genotypes, and the sequence of the PCR products is shown to be SEQ ID No.4, and the female spike and sword leaf length of the 144-strain homozygous CIMMYT8759 genotype F 2 single strain is 2.70+/-2.28 cm; the total 302 strains are heterozygous genotypes, and sequencing of PCR products shows that the sequences of the total 302 strains are SEQ ID No.3 and SEQ ID No.4, and the length of female spike sword leaf of the 302 heterozygous genotype F 2 single strain is 1.10+/-1.28 cm.
Analysis of variance was further performed on the female spike sword leaf length phenotype values for each group (fig. 4). The results show that: the female spike sword leaf length of the homozygous W22 genotype F 2 single plant is extremely lower than that of the homozygous CIMMYT 8759 genotype F 2 single plant, the female spike sword leaf length of the homozygous W22 genotype F 2 single plant is extremely lower than that of the heterozygous genotype F 2 single plant, and the female spike sword leaf length of the heterozygous genotype F 2 single plant is extremely lower than that of the homozygous CIMMYT 8759 genotype F 2 single plant, so that the molecular marker FL3 is related to the female spike sword leaf length of corn, and the method has important breeding application value.
In conclusion, the molecular marker FL3 and qFL3-1 provided by the invention are closely linked, so that the length of the female ear and sword leaf of corn can be rapidly and accurately identified, the application of the locus in the breeding of new varieties of corn can be promoted, and the molecular polymerization breeding of the locus and other excellent character loci can be facilitated. The method provided by the invention can identify and screen the length of the female spike and sword leaf of corn germplasm resources at any stage of corn, has the advantages of simplicity, rapidness, high efficiency and accuracy, and is suitable for large-scale popularization and application.
The present application is described in detail above. It will be apparent to those skilled in the art that the present application can be practiced in a wide range of equivalent parameters, concentrations, and conditions without departing from the spirit and scope of the application and without undue experimentation. While the application has been described with respect to specific embodiments, it will be appreciated that the application may be further modified. In general, this application is intended to cover any variations, uses, or adaptations of the application following, in general, the principles of the application and including such departures from the present disclosure as come within known or customary practice within the art to which the application pertains. The application of some of the basic features may be done in accordance with the scope of the claims that follow.
Claims (8)
1. The application of the substance for detecting the corn female spike and sword leaf length molecular marker in detecting or assisting in detecting the corn female spike and sword leaf length character is that the corn female spike and sword leaf length molecular marker is a DNA fragment shown as SEQ ID No.3 and a DNA fragment shown as SEQ ID No. 4.
2. The use according to claim 1, characterized in that: the substance for detecting the molecular marker of the maize female spike and sword leaf length is a primer pair consisting of two single-stranded DNA shown as SEQ ID No.1 and SEQ ID No.2 in a sequence table.
3. A method for detecting the length traits of female corn ears and swordlike leaves, comprising: the genome DNA of corn to be detected is used as a template, a primer pair consisting of two single-stranded DNA shown as SEQ ID No.1 and SEQ ID No.2 in a sequence table is utilized for PCR amplification, the length of the female spike sword leaf of the homozygous corn with the obtained PCR product sequence of SEQ ID No.3 is shorter or the length of the female spike sword leaf of the homozygous corn with the obtained PCR product sequence of SEQ ID No.4 is shorter or the length of the female spike sword leaf of the homozygous corn with the obtained PCR product sequence of SEQ ID No.3 is shorter or the length of the female spike sword leaf of the heterozygous corn with the obtained PCR product sequences of SEQ ID No.3 and SEQ ID No.4 is shorter or the length of the female spike sword leaf of the heterozygous corn with the obtained PCR product sequence of SEQ ID No.4 is shorter or the candidate.
4. A method for detecting the length traits of female corn ears and swordlike leaves, comprising: the genome DNA of the corn to be detected is used as a template, a primer pair consisting of two single-stranded DNA shown as SEQ ID No.1 and SEQ ID No.2 in a sequence table is used for carrying out PCR amplification, the length of the female spike sword leaf of the homozygous corn with the obtained PCR product size of 178bp is shorter than or candidate shorter than the length of the female spike sword leaf of the homozygous corn with the obtained PCR product size of 363bp, the length of the female spike sword leaf of the homozygous corn with the obtained PCR product size of 178bp is shorter than or candidate shorter than the length of the female spike sword leaf of the heterozygous corn with the obtained PCR product sizes of 178bp and 363bp, and the length of the female spike sword leaf of the heterozygous corn with the obtained PCR product size of 178bp and 363bp is shorter than or candidate shorter than the length of the female spike sword leaf of the homozygous corn with the obtained PCR product size of 363 bp.
5. A method of maize breeding comprising: and (3) taking genome DNA of the corn to be detected as a template, carrying out PCR amplification by using a primer pair consisting of two single-stranded DNA shown as SEQ ID No.1 and SEQ ID No.2 in a sequence table, and selecting the corn to be detected with a PCR product of SEQ ID No.3 as a parent to finish breeding.
6. Use of the substance for detecting the molecular marker of the maize female spike and sword leaf length in claim 1 or 2 for preparing a product for detecting the maize female spike and sword leaf length.
7. The use of the maize female spike and sword leaf length molecular marker of claim 1 or 2 in detecting or assisting in detecting maize female spike and sword leaf length traits.
8. Use of the maize female ear and leaf length molecular marker of claim 1 or 2 in maize breeding.
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