AU2020104105A4 - SNP molecular marker and application of gene GRMZM2G098557 relative to maize ear row number - Google Patents
SNP molecular marker and application of gene GRMZM2G098557 relative to maize ear row number Download PDFInfo
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Abstract
The invention provides SNP molecular markers of gene GRMZM2G098557 relative to maize
ear row number and application thereof, wherein there are two SNP markers, respectively
located at the positions of maize genome Chr4:199959357bp and Chr4:199959358bp, and the
flanking sequence is shown as SEQ ID NO.1. Two SNP molecular markers are significantly
correlated with genes relative to maize ear row number. The row number of maize inbred lines
with the first SNP locus genotype as A/A is higher than that of maize inbred lines with locus
genotype as G/G. The row number of maize inbred lines with the second SNP locus genotype
as G/G is higher than that of maize inbred lines with locus genotype as A/A. The molecular
markers of the invention can be used for maize molecular marker-assisted breeding and
acceleration of the maize high-yield creation and new variety breeding process.
-15
7 ATCTCCATCAGATGTGCCGACTCTCTCAAGCTCATCCACCTTCAGTACCAG
9 ATCTCCATCAGATGTGCCGACTCTGCTCAAGCTCATCCACCTTCAGTACCAG
59 ATCKCATCAGATGTGCCGACTCTGCTCAAGCTCATCCACCTTCAGTACCAG
61 ATCTCCATCAGATGTGCCGACTCTGCTCAAGCTCATCCACCTTCAGTACCAG
85 ATCTCCATCAGATGTGCCGACTCTGCTCAAGCTCATCCACCTTCAGTACCAG
11 ATCTCATCAGATGTGCCGACTCTGCTCAAGCTCATCCACCTTCAGTACCAG
121 ATCTCCATCAGATGTGCCGACTCTGCTCAAGCTCATCCACCTCAGTACCAG
122 ATCTCCA1CAGATGTGCCGACTCTGCTCAAGCTCATCCACCTTCAGTACCAG
155 ATCTCCATCAGATGTGCCGACTCTGCTCAAGCTCATCCACCTTCAGTACCAG
179 ATCTCCATCAGATGTGCC-GACTCTGCTCAAGCTCATCCACCTTCAGTACCAG
187 ATCTCCATCAGATGTGCCGACTCTGCTCAAGCTCATCCACCTTCAGTACCAG
226 ATCTCCA1EAGATGTGCCGACTCTGCTCAAGCTCATCCACCTTCAGTACCAG
263 ATCTCCATCAGATGTGCCGACTCTGCTCAAGCTCATCCACCTTCAGTACCAG
327 ATCTCCATAGATGTGCCGACTCTGCTCAAGCTCATCCACCTTCAGTACCAG
329 ATCTCCATCAGATGTGCCGACTCTGCTCAAGCTCATCCACCTTCAGTACCAG
12 ATCTCCATCAGATGTGCCAGCTCTGCTCAAGCTCATCCACCTCGGTACCAG
14 ATCTCCATCAGATGTGCCAGCTCTGCTCAAGCTCATCCACCTTCGGTACCAG
63 ATCTCCATCAGATGTGCCAGCTCTGCTCAAGCTCATCCACCTTCGGTACCAG
79 ATCTCCATCAGATGTGCCAGCTCTGCTCAAGCTCATCCACCTTCGGTACCAG
84 ATCTCCATCAGATGTGCCAGCTCTGCTCAAGCTCATCCACCTTCGGTACCAG
115 ATCTCCATCAGATGTGCCAGCTCTGCTCAAGCTCATCCACCTTCGGTACCAG
140 ATCTCCATCAGATGTGCCAGCTCTGCTCAAGCTCATCCACCTTCGGTACCAG
144 ATCTCCATCAGATGTGCCAGCTCTGCTCAAGCTCATCCACCTTCGGTACCAG
149 ATCKCATCAGATGTGCCAGCTCTGCTCAAGCTCATCCACCTTCGGTAXCAG
163 ATCTCCATCAGATGTGCCAGCTCTGCTCAAGCTCATCCACCTTCGGTACCAG
214 ATCTCCATCAGATGTGCCAGCTCTGCTCAAGCTCATCCACCTTCGGTACCAG
223 ATCTCCATCAGATGTGCCAGCTCTGCTCAAGCTCAKTCACCTTCGGTACCAG
267 ATCTCCATCAGATGTGCCAGCTCTGCTCAAGCTCATCCACCTTCGGTACCAG
290 ATCTCCATGAGATGTGCCAGCTCTGCTCAAGCTCATCCACCTTCGGTACCAG
311 ATCTCCATCAGATGTGCCAGCTCTGCTCAAGCTCATCCACCTTCGGTACCAG
Figure 1
Description
7 ATCTCCATCAGATGTGCCGACTCTCTCAAGCTCATCCACCTTCAGTACCAG 9 ATCTCCATCAGATGTGCCGACTCTGCTCAAGCTCATCCACCTTCAGTACCAG 59 ATCKCATCAGATGTGCCGACTCTGCTCAAGCTCATCCACCTTCAGTACCAG 61 ATCTCCATCAGATGTGCCGACTCTGCTCAAGCTCATCCACCTTCAGTACCAG ATCTCCATCAGATGTGCCGACTCTGCTCAAGCTCATCCACCTTCAGTACCAG 11 ATCTCATCAGATGTGCCGACTCTGCTCAAGCTCATCCACCTTCAGTACCAG 121 ATCTCCATCAGATGTGCCGACTCTGCTCAAGCTCATCCACCTCAGTACCAG 122 ATCTCCA1CAGATGTGCCGACTCTGCTCAAGCTCATCCACCTTCAGTACCAG 155 ATCTCCATCAGATGTGCCGACTCTGCTCAAGCTCATCCACCTTCAGTACCAG 179 ATCTCCATCAGATGTGCC-GACTCTGCTCAAGCTCATCCACCTTCAGTACCAG 187 ATCTCCATCAGATGTGCCGACTCTGCTCAAGCTCATCCACCTTCAGTACCAG 226 ATCTCCA1EAGATGTGCCGACTCTGCTCAAGCTCATCCACCTTCAGTACCAG 263 ATCTCCATCAGATGTGCCGACTCTGCTCAAGCTCATCCACCTTCAGTACCAG 327 ATCTCCATAGATGTGCCGACTCTGCTCAAGCTCATCCACCTTCAGTACCAG 329 ATCTCCATCAGATGTGCCGACTCTGCTCAAGCTCATCCACCTTCAGTACCAG 12 ATCTCCATCAGATGTGCCAGCTCTGCTCAAGCTCATCCACCTCGGTACCAG 14 ATCTCCATCAGATGTGCCAGCTCTGCTCAAGCTCATCCACCTTCGGTACCAG 63 ATCTCCATCAGATGTGCCAGCTCTGCTCAAGCTCATCCACCTTCGGTACCAG 79 ATCTCCATCAGATGTGCCAGCTCTGCTCAAGCTCATCCACCTTCGGTACCAG 84 ATCTCCATCAGATGTGCCAGCTCTGCTCAAGCTCATCCACCTTCGGTACCAG 115 ATCTCCATCAGATGTGCCAGCTCTGCTCAAGCTCATCCACCTTCGGTACCAG 140 ATCTCCATCAGATGTGCCAGCTCTGCTCAAGCTCATCCACCTTCGGTACCAG 144 ATCTCCATCAGATGTGCCAGCTCTGCTCAAGCTCATCCACCTTCGGTACCAG 149 ATCKCATCAGATGTGCCAGCTCTGCTCAAGCTCATCCACCTTCGGTAXCAG 163 ATCTCCATCAGATGTGCCAGCTCTGCTCAAGCTCATCCACCTTCGGTACCAG 214 ATCTCCATCAGATGTGCCAGCTCTGCTCAAGCTCATCCACCTTCGGTACCAG 223 ATCTCCATCAGATGTGCCAGCTCTGCTCAAGCTCAKTCACCTTCGGTACCAG 267 ATCTCCATCAGATGTGCCAGCTCTGCTCAAGCTCATCCACCTTCGGTACCAG 290 ATCTCCATGAGATGTGCCAGCTCTGCTCAAGCTCATCCACCTTCGGTACCAG 311 ATCTCCATCAGATGTGCCAGCTCTGCTCAAGCTCATCCACCTTCGGTACCAG
Figure 1
SNP molecular marker and application of gene GRMZM2G98557 relative
to maize ear row number
The present invention belongs to the field of molecular genetics, and relates to molecular
markers related to maize ear row number trait, in particular to two SNP markers of
GRMZM2G98557 gene on maize chromosome 4 which are significantly related to
maize ear row number, primer pairs for amplifying the SNP markers and application of
the SNP markers.
Maize is the largest grain crop in China at present, and it is also an important industrial
raw material and energy crop. Continuously increasing the output of maize is an
important guarantee to ensure food security and rapid economic development in China.
With the decrease of cultivated land area in China, it is the most effective way to increase
maize yield by increasing maize yield per unit area. The single yield of maize is mainly
determined by the number of rows per ear, the number of grains per row and the weight
of 100 grains. Therefore, increasing the number of rows per ear is one of the main ways
to increase the yield per unit area of maize.
The research and production practice show that the number of rows per ear is an
important yield component of maize, which has a significant positive correlation with
maize yield. In the process of domestication and genetic improvement of maize, the
number of rows per ear is strongly selected. As an important quantitative trait, the number of rows per ear is controlled by micro-effective polygenic loci, and the development of rows per ear is an important link in the development of maize inflorescence, which is regulated by genes related to inflorescence development. Among the three main components of maize yield, the number of grains per row and 100-grain weight of different varieties are greatly affected by external environmental conditions, and because the number of rows per ear is determined in the process of female ear differentiation, its heritability is high and is less affected by external environmental conditions, which plays an important role in ensuring maize yield. Although some progress has been made in searching maize ear number genes, it is still relatively slow on the whole, and most of the cloned genes related to maize ear number are from mutant research, while the key genes obtained by map-based cloning and with high utility value for breeding practice are still few.
Genome-wide association study (GWAS) is an analytical method based on linkage
disequilibrium (LD) to identify the relationship between phenotypes and genetic markers
in natural populations, and it is an effective way to mine excellent alleles. At present,
with the rapid development of gene sequencing technology and bioinformatics, GWAS
has become one of the most effective methods for identifying the genetic basis of maize
rows per ear. Exploring genes related to the number of rows per ear and developing
functional markers can accelerate the directional genetic improvement of high-yield
maize at the whole genome level.
The first object of the present invention is to provide a set of SNP molecular markers of
the gene GRMZM2G98557 related to the number of rows per ear of maize, and the SNP
markers are significantly related to the number of rows per ear of maize.
The second object of the present invention is to provide a primer pair for amplifying the
SNP molecular marker.
The third object of the present invention is to provide an application of the SNP
molecular marker.
In order to achieve the above purpose, the invention adopts the following technical
scheme.
Collecting 310 maize inbred lines of temperate zone, tropical zone and subtropical zone
in China, the United States, Mexico and so on as the related population of the study.
Combined with the ear row data and SNP genotype data of the associated population,
FarmCPU package in R language is used for GWAS. The results show that the molecular
marker of SNP located at the maize genome Chr4:199959357bp (genome version: Maize
B73RefGen v3) is significantly correlated with the number of rows in the ear of maize,
and is the first molecular marker. Furthermore, it is found from amplification sequencing
that the SNP molecular marker located in maize genome Chr4: 199959358bp (genome
version: Maize B73 RefGenv3) is closely linked with the SNP marker of association
analysis results, which is significantly correlated with the number of rows per ear of
maize, and is the second SNP molecular marker.
The alleles of the first SNP marker are A and G, and there are two homozygous
genotypes of A/A and G/G in the tested inbred lines. Besides, the alleles of the second
SNP molecular marker are A and G, and there are two homozygous genotypes of A/A
and G/G in the tested inbred lines, with a common flanking sequence as shown in SEQ
ID NO.1
The SNP markers are significantly correlated with the number of rows per ear of maize,
and the number of rows per ear of maize inbred lines with genotype A/A at the first SNP
locus is higher than that of maize inbred lines with genotype G/G. The number of rows
per ear of maize inbred lines with the second SNP locus genotype G/G is higher than that
of maize inbred lines with the locus genotype A/A.
Based on the flanking sequences of the significantly associated SNP, the inventor designs
a characteristic primer pair of gene GRMZM2G098557 containing the SNP site for
detecting the maize ear row number, and the sequence is as follows:
Upstream (F): 5'-TGCTGCGGATCAATTCTGGT-3'(SEQ ID NO.2).
Downstream (R): 5'-TACAGACCCATAGCAAGCC-3'(SEQ ID NO.3).
The invention also provides a kit for detecting the SNP molecular marker, which
comprises the above primer pair.
The invention also provides the application of the SNP molecular marker, primer pair or
kit in identifying the gene GRMZM2G98557 related to the number of rows per ear of
maize as well as the application in screening or identifying maize germplasm resources
with the trait of number of rows per ear and its application in improving high yield of
maize.
The invention also provides the application of the SNP molecular marker, primer pair or
kit in breeding high-yield maize.
The invention also provides a method for detecting the number of rows per ear of maize,
which comprises the following steps. Utilizing above primer pair to carry out PCR
amplification on the maize genome DNA to be detected; determining the SNP genotype
of the maize material to be detected according to PCR amplification products; predicting
the number of rows per ear of the maize to be detected. The row number of maize inbred
lines with the first SNP locus genotype as A/A is higher than that of maize inbred lines
with locus genotype as G/G. The row number of maize inbred lines with the second SNP
locus genotype as G/G is higher than that of maize inbred lines with locus genotype as
PCR procedure is pre-denaturation at 94°C for 2 min, degenerating at 98°C for 10 s,
annealing at 59°C for 30s, extending at 68°C for 6 s, with a total of 35 cycles. And then
annealing again at 68°C for 10min.
The inventors use 240 families from the constructed IBM Syn1ODH (B73xMol7)
population as linkage population, and carry out QTL mapping based on the bin marker
genotype data and phenotypic data of maize ear rows. The results show that the above
SNP molecular markers are located in the QTL, which further verifies the importance of
above SNP molecular markers to the number of rows per ear in maize.
In order to further verify the validity of the developed marker, the inventors select 30
families and DNA of two parents (B73 and Mo17) in the linkage population as templates,
and amplify and then sequence by using the above PCR amplification program and
primer pair, and finally verify the locus. The results show that the SNP molecular marker
is successfully typed in 32 maize inbred lines (seen in figure 1). Moreover, the row
number of maize inbred lines with the first SNP locus genotype as A/A is higher than that of maize inbred lines with locus genotype as G/G. The row number of maize inbred lines with the second SNP locus genotype as G/G is higher than that of maize inbred lines with locus genotype as A/A (seen in table 2 and table 3).
The invention utilizes the strategy of combining GWAS with QTL positioning to explain
the internal relationship between the gene and the number of rows per ear of maize, and
find SNP sites with significant function, and the SNP sites. Beside, they are used as
genetic markers for molecular breeding, which is of great significance for accelerating the
high-yield breeding process of maize.
The method disclosed by the invention has the beneficial effects that SNP sites which are
significantly associated with specific traits can be rapidly and accurately detected by
combining GWAS with QTL positioning strategies. Two SNP loci (Maize
B73RefGen_v3:Chr4:199959357, Maize B73RefGen_v3:Chr4:199959358) at the
position of 199959357 bp on chromosome 4 of maize are significantly correlated with the
number of rows per ear, and they can be used as genetic markers for molecular breeding,
which can speed up the breeding process of high-yield maize and have a better
application value.
Figure 1 is the sequencing results of 32 maize inbred lines with amplified SNP sites by
PCR. The box identifies AG as the genotypes of the above two SNP loci of the inbred
lines of maize with high ear row number, while the corresponding locus genotypes of the
inbred lines of maize with low ear row number are GA.
-'7
The following is a further explanation of the technical scheme of the present invention
and the technical effect produced by it in combination with the specific experimental
scheme and the attached figures. The following explanations are only for the purpose of
explaining the invention, but do not restrict the invention in any way. Any exchange or
substitution based on the teaching of the present invention shall be within the protection
scope of the present invention. The methods mentioned in the invention are conventional
methods in this field unless otherwise specified. The reagents used are commercially
available unless otherwise specified.
Embodiment 1 Obtaining SNP molecular marker with significant correlation between
maize gene GRMZM2G98557 and row number per ear trait.
The obtaining method is as follows.
1) Collecting 310 parts of maize inbred lines from China, America, Mexico and other
places in temperate zone, tropical zone and subtropical zone, and constructing the
association population for association mapping. This population is rich in genetic
diversity.
2) Field experiment design. They are planted in Xishuangbanna of Yunnan Province
(experimental base of maize institute of Sichuan Agricultural University), Ya'an of
Sichuan Province (experimental base of maize institute of Sichuan Agricultural
University) and Hongya of Sichuan Province in 2016. The design scheme of field
experiment adopts random block design, with two replicates for each design. Each family
material is planted in two rows, with a length of 4 meters, a row spacing of 0.8 meters,
and a single row of 16 plants, with a planting density of about 50,000 plants per hectare.
Fertilization, watering, weeding and other field management measures are all managed
according to the local normal level. Harvesting after physiological maturity, and the
harvested ears of each inbred line material are used as phenotypic identification samples.
3) Phenotypic identification of related groups. After drying in the sun, 10 maize ears with
the same growth are selected from each inbred line material for phenotypic
determination. Joint analysis of variance shows that the number of rows per ear is
significantly different among environment, genotype and the interaction effect of
environment and genotype (table 1).
Table 1 Joint variance analysis of rows per ear
Source Quadratic Degree of Meanquare Sig Sum Freedom Calibration Model 27201.639 826 32.932 <0.001 Intercept 611384.873 1 611384.873 <0.001 E*G 8783.058 514 17.088 <0.001 E 1440.371 2 720.186 <0.001 G 15143.903 309 49.009 <0.001 Error 6656.174 583 11.417 Gross 759563.280 1410 Calibration Gross 33857.813 1409
4) GWAS. Combining the phenotypic data of maize ear rows in step 3 with high-density
SNP molecular markers, the genome-wide association study is carried out by using the
FarmCPU package in R language, and the allele frequency threshold is set to 0.05. At the
level of P < 1/N (N is the number of SNP markers), the significance of the association
between SNP markers and traits is judged. Results detect an SNP locus significantly
associated with the maize ear rows, with the position of Chr4:199959357 bp, allele of
A/G. Besides, for the SNP locus in selected inbred lines, there are two homozygous
genotypes- A/A and G/G. The sequencing results indicate that an SNP with alleles of G/A in Chr4:199959358 bp position is closely linked to the SNP, wherein, it has two homozygous genotypes G/G and A/A in the selected inbred lines and its flanking sequence is shown as SEQ ID NO. 1.
) QTL mapping verification. Test materials: maize IBM Syn10 DH population,
constructed from the parents B73 and Mo17, containing 280 families and introduced from
Iowa state university. After the investigation of the previous field experiment, 250 lines
of adaptive population and 2 parents are selected as experimental materials.
6) Field experiment design. They are planted in Xishuangbanna of Yunnan Province
(experimental base of maize institute of Sichuan Agricultural University), Chongzhou of
Sichuan Province (experimental base of maize institute of Sichuan Agricultural
University) and Xinxiang of Henan Province (experimental base of Henan Academy of
Agricultural Sciences) in 2015. The design scheme of field experiment adopts random
block design, with two replicates for each design. Each family material is planted in two
rows, with a length of 4 meters, a row spacing of 0.8 meters, and a single row of 16
plants, with a planting density of about 50,000 plants per hectare. Fertilization, watering,
weeding and other field management measures are all managed according to the local
normal level.
7) Phenotypic investigation. Each family in maize IBM Syn10 DH population is
pollinated in an open way, harvested after physiological maturity, and the harvested ears
of each family are used as phenotypic identification samples. After being dried in the sun,
maize ears with the same growth are selected from each family for phenotypic
determination.
8) QTL analysis. Using software QTL Cartographer Version 1.17f, combined with
phenotypic data and 6618 bin marker genotype data, QTL mapping is carried out, in
which a QTL is detected in 185.8 ~ 209.5 Mb on chromosome 4, with a phenotypic
contribution rate of 5.52%. The SNP molecular marker detected in step 4 falls within the
QTL, which further proves the importance of the SNP marker to the number of rows per
ear in maize.
Embodiment 2 Application test of SNP markers of the present invention on the number of
rows per ear of maize.
Selecting 30 pedigree materials with different phenotypes and two parents B73 and Mo17
(the related objects include B73 and Mo17, and the genotype of B73 at two SNP sites is
A/A and G/G, and the genotype of Mo17 is G/G and A/A) from the linkage population,
using its genomic DNA as a template, using the flanking sequences of SNP molecular
marker sites to design specific primers, and the primer sequences are shown in SEQ ID
NO.2 and 3. KOD FX Neo high-fidelity polymerase (Toho (Shanghai) Biotechnology
Co., LTD.) is used for PCR amplification. The procedure is: pre-denaturation at 94°C for
2min, denaturation at 98°C for 10s, annealing for 30s at 59°C, 6s extension at 68°C, with
cycles in total. And then it is extended at 68°C for another 10min. The specific target
band is recovered and purified by gel recovery kit (Omega Bio-Tek), and sequenced by
connecting with the flat-ended cloning vector Peasy-Blunt Cloning Vector (Beijing
Perfect Gold Biotechnology Co.). SnapGene2.3.2 software is used to compare the
sequencing results (seen in figure 1). The results show that this SNP site is successfully
genotyped in 32 selected maize materials to be identified, of which 16 maize materials
with high ear row number have all A/A and G/G genotypes at the two SNP sites, and the other 16 maize materials with low ear row number have all G/G and A/A genotypes at the sites. Moreover, there are significant differences in phenotypic data of genotypes A/A and G/G (table 2 and table 3).
Table 2 Phenotypic data of maize materials to be detected
Number Ear Row Number First SNP Genotype Second SNP Genotype B73 14.29 A G 12 14.83 A G 14 15.10 A G 63 15.70 A G 79 15.40 A G 84 16.40 A G 115 15.27 A G 140 14.90 A G 144 17.50 A G 149 15.80 A G 163 15.12 A G 214 14.97 A G 223 15.80 A G 267 15.07 A G 290 16.90 A G 311 14.90 A G Mol7 10.27 G A 7 13.00 G A 9 11.00 G A 59 12.67 G A 61 12.20 G A 85 11.77 G A 111 12.40 G A 121 12.50 G A 122 10.70 G A 155 12.37 G A 179 12.97 G A 187 11.90 G A 226 12.23 G A 263 12.87 G A 327 11.93 G A 329 11.40 G A
Table 3 t-test results
qaine t-test for the mean equation 95% confidence in Fg Sg Themean Standard difference F Sig. t df (both difrne err The The sides) upper lower limit limit Assuming thatthe 11.97 30 0 3.48562 0.2912 2.89092 4.08033 variances Ear are equal row Assuming 0 0.994 number thatthe variances 11970 29.977 0 3.48562 0.2912 2.8909 4.08035 are unequal
This experiment further proves the accuracy of GWAS and QTL mapping. Therefore, the
above SNP is closely linked with the number of rows per ear in maize. The invention
further confirms that the SNP locus can be used as an effective genetic marker for
molecular marker-assisted selection to improve the ear row number of maize.
Although the present invention has been described in detail by general description,
specific embodiments and experiments, it is obvious to those skilled in the art that some
modifications or improvements can be made on the basis of the present invention.
Therefore, these modifications or improvements made on the basis of not deviating from
the spirit of the invention should belong to the scope of the invention.
SEQ ID NO.1: atgtcagatg ttaaatactg gtaccgaagg tggatgagct tgagcagagc nngcacatct 60 gatggagatg ttttagacgt aatcacagcc cggggggata 100 2020104105
SEQ ID NO.2: 5'-tgctgcggatcaattctggt -3' 20
SEQ ID NO.3: 5'-tacagacccatacgcaagcc-3' 20
Claims (10)
1. A set of SNP molecular marker of gene GRMZM2G098557 relative to maize ear row
number is characterized by comprising a first SNP marker and a second SNP marker.
Wherein, the first SNP marker is located at the maize genome Chr4:199959357bp; the
second SNP marker is located at the maize genome Chr4:199959358bp.
The alleles of the first and second SNP markers are A and G.
2. The SNP molecular marker according to claim 1 is characterized in that the flanking
sequence of the allele is shown in SEQ ID NO.1
3. The SNP molecular marker according to claim 1 or 2 is characterized in that the SNP
molecular marker is significantly correlated with maize ear row number. Wherein, the
row number of maize inbred lines with the first SNP locus genotype as A/A is higher than
that of maize inbred lines with locus genotype as G/G. The row number of maize inbred
lines with the second SNP locus genotype as G/G is higher than that of maize inbred lines
with locus genotype as A/A.
4. A primer pair for detecting a set of SNP markers according to any one of claims 1-3, is
characterized in that the primer pair has a nucleotide sequence shown in SEQ ID NO.2-3.
5. The kit for detecting a set of SNP molecular markers according to any one of claims1
3, is characterized by comprising the primer pair in claim 4.
6. Application of a set of SNP molecular marker based on any one of claims 1-3, a primer
pair based on claim 4 or a kit in claim 5 in identifying the gene GRMZM2G098557
related to the number of rows per ear in maize.
7. Application of a set of SNP molecular marker based on any one of claims 1-3, a primer
pair based on claim 4 or a kit in claim 5 in screening or identifying maize germplasm
resources with the number of rows per ear.
8. Application of a set of SNP molecular marker based on any one of claims 1-3, a primer
pair based on claim 4 or a kit in claim 5 in improving high yield of maize.
9. A method for detecting the number of rows per ear of maize is characterized in that the
genomic DNA of the maize material to be detected is amplified by PCR with the primer
pair of claim 4, and the genotype of each SNP marker in the set of maize to be detected is
determined according to the PCR amplification product. And then predicting the number
of rows per ear of the maize to be detected based on each genotype of the maize SNP
markers to be detected.
10. The method according to claim 9 is characterized in that the PCR procedure is as
follows: pre-denaturation at 94°C for 2 min, degenerate at 98°C for 10 s, annealing at
59°C for 30s, extending at 68°C for 6 s with a total of 35 cycles. And then extending
again at 68°C for 10min.
Figure 1
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115927736A (en) * | 2023-01-30 | 2023-04-07 | 河北师范大学 | Wheat spikelet number related SNP (single nucleotide polymorphism) site and application thereof |
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2020
- 2020-12-15 AU AU2020104105A patent/AU2020104105A4/en not_active Ceased
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115927736A (en) * | 2023-01-30 | 2023-04-07 | 河北师范大学 | Wheat spikelet number related SNP (single nucleotide polymorphism) site and application thereof |
| CN115927736B (en) * | 2023-01-30 | 2023-06-30 | 河北师范大学 | SNP locus related to wheat spike number per spike and application thereof |
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