CN115820897A - Molecular marker closely linked with length of maize female ear and sword leaf and application thereof - Google Patents
Molecular marker closely linked with length of maize female ear and sword leaf and application thereof Download PDFInfo
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Abstract
The invention discloses a molecular marker closely linked with the length of corn ear and sword leaf and application thereof. The molecular marker closely linked with the length of the corn ear and the sword leaf is a DNA fragment shown in SEQ ID No.3 or a DNA fragment shown in SEQ ID No.4, and can be detected by using a primer pair consisting of two single-stranded DNAs shown in SEQ ID No.1 and SEQ ID No.2 in a sequence table. Experiments prove that the molecular marker closely linked with the length of the maize female ear sword leaf is related to the length of the maize female ear sword leaf, the molecular marker can be used for successfully identifying the length of the maize female ear sword leaf, and the molecular marker has the advantages of simplicity, convenience, rapidness, high efficiency, accuracy, good repeatability and high specificity, can be used for maize molecular marker assisted breeding, breeds new maize varieties with excellent comprehensive properties, greatly saves breeding cost and improves breeding efficiency.
Description
Technical Field
The invention relates to a molecular marker closely linked with the length of corn ear and sword leaf and application thereof in the field of biotechnology.
Background
Corn (Zea mays L.) is a crop which is used for both food, feed and economy, and is the only crop which is steadily increased in sowing area and yield in China. In recent years, global economy has been rapidly developed, and the demand for corn is expected to increase in future industrial production and life. In 2021, the planting area of the corn in China is 4332 ten thousand hectares, 206 ten thousand hectares are increased compared with the planting area in the last year, the yield is up to 27255 ten thousand tons, and the yield accounts for about 39.91 percent of the total yield of the grain in China. Therefore, the corn production has an important strategic position in guaranteeing the food safety in China.
The sword leaf is further extension of the top end of the bract of the female ear of the corn, is an organ degenerated in the long-term evolution process of the corn, and is one of important characters for identifying corn varieties and germplasm sources. The difference of the lengths of the flag leaves of the corns from different germplasm sources is large, most of the sweet corns in temperate zone have long flag leaves, and most of the tropical corns have no flag leaves or have short flag leaves. The sword leaf is used as an important component of the corn ear and plays an important role in the growth, development and yield formation of the corn ear. Therefore, further analyzing the genetic basis of the maize female ear and sword leaf length and developing a molecular marker closely linked with the Quantitative Trait Locus (QTL) of the female ear and sword leaf length have important significance for breeding a new excellent maize variety with proper female ear and sword leaf length trait and great market economic value.
The molecular markers have the advantages of large quantity, no influence of environmental conditions, development period, expression regulation and other factors on detection, capability of providing complete and rich genetic information and the like, and are widely applied to the aspects of germplasm resource identification, QTL positioning, molecular marker-assisted selection and the like. An Insertion-Deletion (InDel) marker is one of the commonly used molecular markers based on the difference of DNA levels, and specifically refers to the difference between two materials, wherein, relative to one material, a certain number of nucleotide insertions or deletions exist in certain sites of the genome of the other material, and PCR primers for amplifying the Insertion-Deletion sites are designed according to the Insertion-Deletion sites. By utilizing the InDel marker which is tightly linked with the target gene, linkage drag is reduced, favorable genes are aggregated, the breeding process is accelerated, and the selection efficiency and effect can be effectively improved through auxiliary backcross selection, auxiliary pedigree selection and even whole genome selection.
At present, although research reports show that the QTL for controlling the length of the xiphoid leaves of the maize ears, no molecular marker which is closely linked with the target QTL is developed and applied for patent, and no patent report related to the length of the xiphoid leaves of the maize ears is reported in a qFL3-1 segment.
Disclosure of Invention
The invention aims to solve the technical problem of how to detect the length character of the maize female ear and the sword leaf.
In order to solve the technical problems, the invention firstly provides the application of a substance for detecting the molecular marker of the length of the maize female ear and sword leaf in the detection or the auxiliary detection of the length character of the maize female ear and sword leaf, wherein the molecular marker of the length of the maize female ear and sword leaf 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 maize ear and sword leaf length can be a primer pair consisting of two single-stranded DNAs 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 character of the maize female ear and the sword leaf, which comprises the following steps: taking genome DNA of a corn to be detected as a template, and carrying out PCR amplification 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, wherein the length of the female tassel and sword leaf of the homozygous corn with the PCR product sequence of SEQ ID No.3 is shorter or candidate shorter than that of the female tassel and sword leaf of the homozygous corn with the PCR product sequence of SEQ ID No.4, the length of the female tassel and sword leaf of the homozygous corn with the PCR product sequence of SEQ ID No.3 is shorter or candidate shorter than that of the female tassel and sword leaf of the heterozygous corn with the PCR product sequences of SEQ ID No.3 and SEQ ID No.4, and the length of the female tassel and sword leaf of the heterozygous corn with the PCR product sequences of SEQ ID No.3 and SEQ ID No.4 is shorter or candidate shorter than that of the female tassel and sword leaf of the homozygous corn with the PCR product sequence of SEQ ID No.4.
The invention also provides a method for detecting the length character of the maize female ear and the sword leaf, which comprises the following steps: using genome DNA of corn to be detected as a template, and utilizing a primer pair consisting of two single-stranded DNAs shown as SEQ ID No.1 and SEQ ID No.2 in a sequence table to perform PCR amplification, wherein the length of the female tassel and sword leaf of the homozygous corn with the PCR product size of 178bp is shorter than or is candidate to be shorter than the length of the female tassel and sword leaf of the homozygous corn with the PCR product size of 363bp, the length of the female tassel and sword leaf of the homozygous corn with the PCR product size of 178bp is shorter than or is candidate to be shorter than the length of the female tassel and sword leaf of the heterozygous corn with the PCR products sizes of 178bp and 363bp, and the length of the female tassel and sword leaf of the heterozygous corn with the PCR products sizes of 178bp and 363bp is shorter than or is candidate to be shorter than the length of the homozygous corn with the PCR product size of 363 bp.
The invention also provides a method for breeding corn, which comprises the following steps: taking the genome DNA of the corn to be tested as a template, carrying out PCR amplification 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, and selecting the corn to be tested with the PCR product of SEQ ID No.3 as a parent to finish breeding.
In the above, the reaction system for performing PCR amplification using the 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: 1 mu L of single-stranded DNA shown in SEQ ID No.1 with the concentration of 10 mu mo 1/L; 1 mu L of single-stranded DNA shown in SEQ ID No.2 with the concentration of 10 mu mo 1/L; 1 μ L of genomic DNA at a concentration of 100 ng/. Mu.L; 5 μ L of 2 XTAQQ PCR StarMix with Loading Dye (Biotech, inc., kangcheng Chengcheng, beijing, cat. No. A012-01); 2 μ L ddH 2 O。
The reaction conditions for PCR amplification by using a primer pair consisting of two single-stranded DNAs shown by SEQ ID No.1 and SEQ ID No.2 in the sequence table may be: pre-denaturation at 95 ℃ for 10min; denaturation at 95 ℃ for 45s, annealing at 57 ℃ for 45s, extension at 72 ℃ for 35s,35 cycles; extending for 10min at 72 ℃; storing at 4 ℃.
The application of the substance for detecting the molecular marker of the length of the maize female ear xiphoid leaf in preparing products for detecting the length characters of the maize female ear xiphoid leaf also belongs to the protection scope of the invention.
The application of the molecular marker for the length of the maize female ear and the sword leaf in detecting or assisting in detecting the length character of the maize female ear and the sword leaf also belongs to the protection scope of the invention.
The application of the molecular marker for the length of the maize female ear and the sword leaf in maize breeding also belongs to the protection scope of the invention.
In the present invention, the xiphoid leaf length of the female ear may be the length of the maize that extends further out from the top of the maize ear bract.
The molecular marker for the length of the female ear and the sword leaf of the corn is related to the length of the female ear and the sword leaf of the corn, can be used for successfully identifying the length of the female ear and the sword leaf of the corn, has the advantages of simplicity, convenience, rapidness, high efficiency and accuracy, has good repeatability and high specificity, can be used for the molecular marker-assisted breeding of the corn, breeds a new corn variety with excellent comprehensive properties, greatly saves the breeding cost and improves the breeding efficiency.
The present invention is described in further detail below with reference to specific embodiments, and the examples are given only for illustrating the present invention and not for limiting the scope of the present invention. The examples provided below serve as a guide for further modifications by a person skilled in the art and do not constitute a limitation of the invention in any way.
Drawings
FIG. 1 shows the alignment of the amplified sequence of the molecular marker qFL3-1 of the present invention in parents.
FIG. 2 is an electropherogram of PCR amplification product of molecular marker qFL3-1 of the present invention in parents. Wherein W is the amplification band type of a maize inbred line W22, C is the amplification band type of a maize wild relative species CIMMYT8759, and the sizes of bands of a Marker are 100bp, 250bp, 500bp, 750bp, 1000bp, 1500bp and 2000bp from bottom to top in sequence.
FIG. 3 shows the molecular marker qFL3-1 of the present invention at F 2 Electropherograms of PCR amplification products in the population. Wherein, W is the amplification band type of the homozygous W22 genotype, C is the amplification band type of the homozygous CIMMYT8759 genotype, H is the amplification band type of the heterozygous genotype, and the sizes of the bands of the Marker from bottom to top are 100bp, 250bp, 500bp, 750bp, 1000bp, 1500bp and 2000bp in sequence.
FIG. 4 shows the molecular marker qFL3-1 of the present invention at F 2 Single-marker analysis results of the length traits of the pistil sword leaves in the population. NIL _ W22 indicates homozygous W22 genotype, het indicates heterozygous genotype, NIL _ CIMMYT8759 indicates homozygous CIMMYT8759 genotype, and x indicates very significant difference (P)<0.01)。
Detailed Description
The experimental procedures in the following examples, unless otherwise indicated, are conventional and are carried out according to the techniques or conditions described in the literature in the field or according to the instructions of the products. Materials, reagents, instruments and the like used in the following examples are commercially available unless otherwise specified. The quantitative tests in the following examples, all set up three replicates 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 relative species CIMMYT8759 of the following examples are described in the literature (Identification and fine mapping of the qualitative tragic locus for the number of the molecular bundle in the medium, JIntegrplant biol.2016Jan;58 (1): 81-90. Doi.
MR0242 in the following example is a product of the American corn germplasm storage Center (Maize Genetics Cooperation storage Center) with the website as: weww. Maizecoop. Chopper. Uiuc. Edu, the biological material is also available to the public from the applicant, and is only used for repeating the relevant experiments of the present invention, and is not used for other purposes. MR0242 is introgression line material derived from maize inbred line W22 and maize wild relative species CIMMYT8759 by crossing, backcrossing and selfing.
Example 1: molecular marker closely linked with length of maize female ear and sword leaf
The invention provides a molecular marker qFL3-1 (marked as a corn ear and sword leaf length molecular marker) for identifying or assisting in identifying the length of corn ear and sword leaves, wherein the molecular marker is a DNA fragment obtained by performing PCR amplification on A1 by using a primer with 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 pair A1 has the following sequences:
forward amplification primers FL3-F:5 'GCCCCTGTCGATGTTCAAC-3', as shown in SEQ ID No. 1;
reverse amplification primers FL3-R:5 'AGAATTCCAGTTGCTTAAGAGGC-3', as shown in SEQ ID No. 2;
taking genome DNA of a maize inbred line W22 with short xiphoid leaf length and a maize wild kindred species CIMMYT8759 with long xiphoid leaf length as templates, carrying out PCR amplification by using a forward amplification primer shown in SEQ ID No.1 and a reverse amplification primer shown in SEQ ID No.2, and detecting the sequence of the obtained PCR product.
Wherein, the reaction system for 10 μ L PCR amplification is as follows:
(1) 1 mu L of forward amplification primer shown in SEQ ID No.1 with the concentration of 10 mu mo 1/L;
(2) 1 mu L of reverse amplification primer shown in SEQ ID No.2 with the concentration of 10 mu mo 1/L;
(3) 1 μ L of DNA template with a concentration of 100 ng/. Mu.L;
(4) 5 μ L of 2 XTAQQ PCR StarMix with Loading Dye (Biotech, inc., kangcheng Chengcheng, beijing, cat. No. A012-01);
(5)2μL ddH 2 O。
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, extension at 72 ℃ for 35s,35 cycles;
(3) Extending for 10min at 72 ℃;
(4) Storing at 4 ℃.
The PCR instrument model: eppendorf Mastercycler nexus.
Carrying out electrophoretic separation on the PCR amplification product in 2.0% agarose gel (each 100mL of gel solution contains 2.0g of agarose), and carrying out sequencing analysis, wherein the result shows that the PCR amplification is carried out by taking the genome DNA of the maize inbred line W22 as a template, the molecular weight of the PCR amplification product is 178bp, and the nucleotide sequence is shown as SEQ ID No. 3; and (3) carrying out PCR amplification by using the genome DNA of the corn wild relative species CIMMYT8759 as a template, wherein the molecular weight of a PCR amplification product is 363bp, and the nucleotide sequence of the PCR amplification product is shown as SEQ ID No.4.
SEQ ID No.3：
GCCCCTGCGATGTTCAACATACTGCTCATAGGCATAACTTGAGGGTGTGTGGGGCTTATCATAGCACCATTGGATGGATCTCAAAATTTAAAATTTGGCGCACATATTGCATCATGGAATACTATATTACCGTGTTCAGCATCTATACTCTTTGTGCCTCTTAAGCAACTGGAATTCT。
SEQ ID No.4：
GCCCCTGCGATGTTCAACATACTGCTCATAGGCATAACTTGAGGGTGTGTGGGGCTTATCATAGCACCATTGGATGCAGGGGCGAAGCCAGCATTTAAGATTGAGAGGGACAAATTAGATTGAGGGGGGCTGTTAAGAGGTATTTTACATTATTTATATGGTGATTAGCTAAAAAAATTAGTAGCTTCTATGGAATTTGTAGAAGATTAGGGGGGACATTGCCCCCCTGTGCCCCTCCCTAGAATCGCCCCTGATTGGATGGATCTCAAAATTTAAAATTTGGCACACATATTGCATCATGGAATACTATATTACCGTGTTCAGCATCTATACTCTTTGTGCCTCTTAAGCAACTGGAATTCT。
Wherein, the amplified banding pattern of the maize inbred line W22 is an allele for shortening the length of the female ear and the sword leaf. Therefore, if the molecular weight of the PCR amplification product of the corn sample to be detected is 178bp, the corn sample to be detected contains the allele for shortening the length of the pistachio sword-like leaves of the corn; if the molecular weight of the PCR amplification 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 ear and the sword leaf of the corn.
The result of alignment of SEQ ID No.3 and SEQ ID No.4 is shown in FIG. 1. The electrophoresis results of the PCR amplification products of the maize inbred line W22 and the maize wild relative species CIMMYT8759 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 containing 866 families 2 S 3 Introgression line population
Using a maize inbred line W22 as a receptor parent and a maize wild relative species CIMMYT8759 as a donor parent, and obtaining BC containing 866 families by hybridizing 1 generation, backcrossing 2 generation and inbreeding 3 generation 2 S 3 A population of introgression lines.
Step 2: field planting and phenotypic determination of introgression line populations
The crops are planted in 2019 spring in a regional test station of national crop varieties of Liuyang city (28.2 degrees N,113.6 degrees E) in Hunan provinceBC 2 S 3 A population of introgression lines. The field experiment adopts an amplification type incomplete random block design. Each cell is planted with 2 rows, each row is planted with 15 plants, and the plant spacing is 25cm. 2 families are planted on each ridge. The ridge height is 15cm, the ridge width is 70cm, and the furrow width is 30cm.
The tassel length phenotype was measured 10 days after dusting and 10 individuals were investigated consecutively for each family material starting from strain 2. The length of the female ear sword-like leaf is the length of the corn which extends further outwards from the top of the corn female ear bract.
And step 3: QTL location analysis
And carrying out QTL positioning analysis by utilizing a multi-QTL model of R/QTL. Firstly, carrying out QTL simple interval positioning analysis by using Haley-Knott regression, and determining the LOD threshold (alpha = 0.05) of the xiphoid leaf length QTL by adopting a method of 10000 times of replacement test. And carrying out multi-QTL model fitting on the QTL model obtained by simple interval positioning, and optimizing the position of each QTL by utilizing a R/QTL refinqtll command. And further detecting whether other QTL (quantitative trait loci) of the significant improvement model exist in the genome by using the addqtl command, if the new QTL is detected, re-fitting the multi-QTL model and optimizing the QTL position, and repeating the process until the new QTL cannot be detected. And finally, calculating the total phenotypic variation of all QTL interpretations and the additive effect and phenotypic contribution rate of the single QTL by using a fitqtl command.
QTL positioning result analysis: a total of 9 QTLs controlling the length of the maize ear and the xiphoid leaves were detected, wherein a 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 the interval from 132255901bp to 151202327bp on a maize 3 rd chromosome.
And 4, step 4: development and Synthesis of molecular marker FL3
qFL3-1 was searched for the physical interval from 132255901bp to 151202327bp of chromosome 3 by using online primer design software primer3 (https:// primer3.Ut. Ee /), and a forward amplification primer FL3-F and a reverse amplification primer FL3-R were designed, the primers were synthesized by Beijing optimak Biotechnology Ltd, and the nucleotide sequences were as follows:
forward amplification primers FL3-F:5 'GCCCCTGTCGATGTTCAAC-3', as shown in SEQ ID No. 1;
reverse amplification primers FL3-R:5 'AGAATTCCAGTTGCTTAAGAGGC-3', as shown in SEQ ID No. 2.
Example 3: application of molecular marker FL3
Starting with introgression line MR0242 heterozygous only for the qFL3-1 segment and homozygous at other sites in the genome, self-pollinated to produce an F segregating only for the qFL3-1 segment 2 And (4) a group. So as to contain 584 individuals of F 2 The population is used as a material to verify the molecular marker FL3 obtained by the invention 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: f 2 Determination of the length of the Sword leaf of the population
F is determined as in example 2 2 The length of the female tassel and sword leaves of the plant group.
Step 2: and (3) extracting the DNA of the corn leaves by adopting a CTAB method.
And step 3: PCR amplification
The reaction system for PCR amplification is 10. Mu.L, and comprises:
(1) 1 mu L of forward amplification primer shown in SEQ ID No.1 with the concentration of 10 mu mo 1/L;
(2) 1 mu L of reverse amplification primer shown in SEQ ID No.2 with the concentration of 10 mu mo 1/L;
(3) 1 μ L of DNA template at a concentration of 100 ng/. Mu.L;
(4) 5 μ L of 2 XTAQQ PCR StarMix with Loading Dye (Biotech, inc., kangcheng Chengcheng, beijing, cat. No. A012-01);
(5)2μL ddH 2 O。
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, extension at 72 ℃ for 35s,35 cycles;
(3) Extending for 10min at 72 ℃;
(4) Storing at 4 ℃.
The PCR instrument model: eppendorf Mastercycler nexus.
And 4, step 4: electrophoresis
Molecular marker FL3 in part F 2 The electrophoretogram of the PCR amplified product of the individual strain is shown in FIG. 3.
And 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 amplification product of the corn sample to be detected only has a 178bp strip, the corn to be detected is homozygous W22 genotype (namely the genotype is the same as the genotype of the corn inbred line W22); if the PCR amplification product of the corn sample to be detected only has a 363bp strip, the corn to be detected is homozygous CIMMYT8759 genotype (namely the genotype is the same as that of the wild relative species CIMMYT8759 of the corn); if the PCR amplification 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 in a heterozygous genotype.
F 2 138 strains in a single plant are homozygous W22 genotype, and the sequencing of PCR products shows that the sequences of the W22 genotype are all SEQ ID No.3 and 138 strains are homozygous F22 genotype 2 The length of the female ear and the sword leaf of the single plant is 0.43 +/-0.79 cm; 144 strains are homozygous CIMMYT8759 genotypes, the sequencing of PCR products shows that the sequences of the PCR products are SEQ ID No.4, and 144 strains are homozygous CIMMYT8759 genotypes F 2 The length of the female ear and the sword leaf of the single plant is 2.70 +/-2.28 cm; the total 302 strains are heterozygous genotypes, the sequencing of PCR products shows that the sequences are SEQ ID No.3 and SEQ ID No.4, the 302 strains are heterozygous genotypes F 2 The length of the female tassel and the sword leaf of the single plant is 1.10 +/-1.28 cm.
The anorthose length phenotype values of the ears of each group were further analyzed for variance (fig. 4). The results show that: homozygous W22 genotype F 2 The length of the female ear and sword leaf of the single plant is remarkably lower than that of the homozygous CIMMYT8759 genotype F 2 Individual, homozygous W22 genotype F 2 The length of the female ear and the sword leaf of the single plant is extremely obviously lower than that of the heterozygous genotype F 2 Individual, heterozygous genotype F 2 The length of the female ear and sword leaf of the single plant is remarkably lower than that of the homozygous CIMMYT8759 genotype F 2 The single plant shows that the molecular marker FL3 is related to the length of the female ear and the sword leaf of the corn, and has important breeding application value.
In conclusion, the molecular marker FL3 provided by the invention is closely linked with qFL3-1, so that the length of the xiphoid leaves of the maize ear can be quickly and accurately identified, the application of the locus in breeding of new maize varieties can be promoted, and the molecular polymerization breeding of the locus and other excellent character loci is facilitated. By the method provided by the invention, the length of the female tassel and the sword leaf of the corn germplasm resource can be identified and screened at any stage of the corn, and the method has the advantages of simplicity, convenience, rapidness, high efficiency and accuracy and is suitable for large-scale popularization and application.
The present invention has been described in detail above. It will be apparent to those skilled in the art that the invention can be practiced within a wide range of equivalent parameters, concentrations, and conditions without departing from the spirit and scope of the invention and without undue experimentation. While the invention has been described with reference to specific embodiments, it will be appreciated that the invention can be further modified. In general, this application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the invention and including such departures from the present disclosure as come within known or customary practice within the art to which the invention pertains. The use of some of the essential features is possible within the scope of the claims attached below.
Claims (8)
1. The application of the substance for detecting the molecular marker of the length of the maize female ear and sword leaf in detecting or assisting in detecting the length character of the maize female ear and sword leaf is disclosed, wherein the molecular marker of the length of the maize female ear and sword leaf is a DNA fragment shown in SEQ ID No.3 and a DNA fragment shown in SEQ ID No.4.
2. Use according to claim 1, characterized in that: the substance for detecting the molecular marker of the maize female ear and sword leaf length is a primer pair consisting of two single-stranded DNAs shown as SEQ ID No.1 and SEQ ID No.2 in a sequence table.
3. The method for detecting the length character of the maize female ear and the sword leaf comprises the following steps: taking genome DNA of a corn to be detected as a template, and carrying out PCR amplification 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, wherein the length of the female tassel and sword leaf of the homozygous corn with the PCR product sequence of SEQ ID No.3 is shorter or candidate shorter than that of the female tassel and sword leaf of the homozygous corn with the PCR product sequence of SEQ ID No.4, the length of the female tassel and sword leaf of the homozygous corn with the PCR product sequence of SEQ ID No.3 is shorter or candidate shorter than that of the female tassel and sword leaf of the heterozygous corn with the PCR product sequences of SEQ ID No.3 and SEQ ID No.4, and the length of the female tassel and sword leaf of the heterozygous corn with the PCR product sequences of SEQ ID No.3 and SEQ ID No.4 is shorter or candidate shorter than that of the female tassel and sword leaf of the homozygous corn with the PCR product sequence of SEQ ID No.4.
4. The method for detecting the length character of the maize female ear and the sword leaf comprises the following steps: using genome DNA of corn to be detected as a template, and utilizing a primer pair consisting of two single-stranded DNAs shown as SEQ ID No.1 and SEQ ID No.2 in a sequence table to perform PCR amplification, wherein the length of the female tassel and sword leaf of the homozygous corn with the PCR product size of 178bp is shorter than or is candidate to be shorter than the length of the female tassel and sword leaf of the homozygous corn with the PCR product size of 363bp, the length of the female tassel and sword leaf of the homozygous corn with the PCR product size of 178bp is shorter than or is candidate to be shorter than the length of the female tassel and sword leaf of the heterozygous corn with the PCR products sizes of 178bp and 363bp, and the length of the female tassel and sword leaf of the heterozygous corn with the PCR products sizes of 178bp and 363bp is shorter than or is candidate to be shorter than the length of the homozygous corn with the PCR product size of 363 bp.
5. A method of maize breeding comprising: taking the genome DNA of the corn to be tested as a template, carrying out PCR amplification 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, and selecting the corn to be tested 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 maize female ear and sword leaf length in claim 1 or 2 for preparing a product for detecting the trait of maize female ear and sword leaf length.
7. Use of the molecular marker for maize female ear/sword leaf length according to claim 1 or 2 for detecting or assisting in detecting the trait of maize female ear/sword leaf length.
8. The use of the maize female ear/sword leaf length molecular markers of claim 1 or 2 in maize breeding.
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