CN116621961A - Gene ZmAPC4 for regulating starch content in corn kernel, expression product, SNP marker, excellent haplotype and application - Google Patents

Gene ZmAPC4 for regulating starch content in corn kernel, expression product, SNP marker, excellent haplotype and application Download PDF

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CN116621961A
CN116621961A CN202310642357.9A CN202310642357A CN116621961A CN 116621961 A CN116621961 A CN 116621961A CN 202310642357 A CN202310642357 A CN 202310642357A CN 116621961 A CN116621961 A CN 116621961A
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zmapc4
gene
starch content
corn
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张雪海
汤继华
段海洋
李建新
熊雪航
付志远
丁冬
陈晓阳
李卫华
王洪秋
孙莉
孙岩
徐书豪
薛政杰
琚小龙
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Henan Agricultural University
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Abstract

The invention relates to a gene for regulating and controlling starch content in corn grainsZmAPC4Expression products, SNP markers, excellent haplotypes and application thereof, and belongs to the technical field of genetic engineering breeding. The geneZmAPC4The nucleotide sequence of (2) is shown as SEQ ID NO. 1. The geneZmAPC4The amino acid sequence of the encoded protein is shown as SEQ ID NO. 2. Gene for regulating starch content in corn kernelZmAPC4The haplotype comprises 4 types, and the nucleotide sequences of the haplotypes are shown as SEQ ID NO.3, SEQ ID NO.4, SEQ ID NO.5 and SEQ ID NO. 6. Wherein SEQ ID NO.6 is an excellent haplotype. The invention relates to a regulating gene for starch content in corn kernelZmAPC4Is applied to corn cultivationThe practical method can be popularized and applied to breeding of other related genes.

Description

Gene ZmAPC4 for regulating starch content in corn kernel, expression product, SNP marker, excellent haplotype and application
Technical Field
The invention relates to the technical field of genetic engineering breeding, in particular to a gene ZmAPC4 for regulating and controlling starch content in corn kernels, an expression product, SNP markers, excellent haplotypes and application.
Background
Starch plays an important role in human social activities as an energy source. Grains containing high amounts of starch are useful in the manufacture of food, animal feed, biofuels and other products. The global grain yield in 2022 reaches 28 hundred million tons, and the starch acquisition becomes simple and easy. However, grain safety is still facing tremendous pressure due to the rapid growth of the population and the deterioration of the ecological environment. Currently, increasing the yield per unit area has reached a bottleneck, and further increases in cultivated land area to meet the growing population demand are not feasible. Thus, it is necessary to increase crop dry matter accumulation to produce more starch. The starch content in the corn kernels is about 70%, and increasing the starch content in the corn kernels on the premise of keeping the yield stable is one of the main targets of corn breeding at present. In addition, the research on the genetic mechanism of the corn starch content can be further developed, so that theoretical basis and excellent germplasm resources can be provided for breeding new varieties of corn with high starch content.
Disclosure of Invention
In order to improve the starch content in corn kernels, the invention provides a gene ZmAPC4 for regulating the starch content in corn kernels, an expression product, an SNP marker, an excellent haplotype and application, and the method for applying the starch content regulating gene ZmAPC4 in corn kernels to corn breeding practice can be popularized and applied to breeding application of other related genes.
In order to achieve the above purpose, the invention adopts the following technical scheme:
a gene ZmAPC4 for regulating and controlling starch content in corn kernels is shown in SEQ ID NO. 1.
The gene ZmAPC4 expression product for regulating and controlling the starch content in corn grains, wherein the gene ZmAPC4 expression product is protein coded by the gene ZmAPC4, and the amino acid sequence of the gene ZmAPC4 expression product is shown as SEQ ID NO. 2.
A gene ZmAPC4 SNP marker for regulating and controlling starch content in corn grains is used for amplifying a primer pair containing a ZmAPC4 intramolecular specific SNP marker, wherein an upstream primer sequence is shown as SEQ ID NO.7, and a downstream primer sequence is shown as SEQ ID NO. 8.
The gene ZmAPC4 haplotype for regulating and controlling the starch content in corn kernel includes 4 kinds, and the nucleotide sequences are shown in SEQ ID No.3, SEQ ID No.4, SEQ ID No.5 and SEQ ID No. 6. Wherein SEQ ID NO.6 is an excellent haplotype.
A molecular marker for distinguishing starch content in corn grains has an upstream primer sequence shown in SEQ ID NO.9 and a downstream primer sequence shown in SEQ ID NO. 10.
An application of a gene ZmAPC4 for regulating and controlling starch content in corn kernels in corn genetic breeding.
An application of a gene ZmAPC4 excellent haplotype for regulating and controlling starch content in corn kernels in corn genetic breeding.
Compared with the prior art, the invention has the beneficial effects that:
1. the excellent haplotype of the starch content regulating gene ZmAPC4 in the corn kernel is suitable for identifying corn germplasm resources with high starch content, and the molecular marker developed based on the ZmAPC4 can be used for distinguishing corn inbred lines with different starch contents, so that the line selection efficiency in the corn breeding process is improved. Has higher practical value and provides reference for cultivating new corn varieties with high starch content.
2. The nucleotide sequence of the ZmAPC4 gene obtained by screening is shown as SEQ ID NO.1, and the encoded protein sequence is shown as SEQ ID NO. 2. Based on the ZmAPC4 intramolecular SNP marker, a batch of excellent germplasm resources with higher starch content are screened, and in addition, dCAPS markers are also developed for distinguishing corn inbred lines with different starch contents. These results indicate that the ZmAPC4 gene can be used in corn genetic breeding and has positive significance for improving the starch content in corn kernels.
Drawings
In order to more clearly illustrate the embodiments of the invention or the technical solutions in the prior art, the drawings that are required in the embodiments or the description of the prior art will be briefly described, it being obvious that the drawings in the following description are only some embodiments of the invention, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a Q-Q plot and Manhattan plot generated from a genome-wide correlation analysis of starch content in corn kernels using a Q model;
wherein, the graph A represents a Q-Q graph generated by carrying out whole genome correlation analysis on starch content in corn kernels;
panel B shows a Manhattan plot generated from a whole genome correlation analysis of starch content in corn kernels.
FIG. 2 shows the results of GO (Gene Ontology) enrichment analysis of expressed genes in corn kernels screened using whole genome correlation analysis.
FIG. 3 shows the analysis of the differences between the different haplotypes and haplotype combinations of the starch content controlling gene ZmAPC4 in corn kernels, wherein, the abscissa Hap1, hap2, hap3 and Hap4 respectively represent the different haplotypes.
FIG. 4 is a differential analysis of starch content in corn inbred grain carrying ZmAPC4 intramolecular specific SNP marker (TT or GG).
FIG. 5 is a photograph of UV imaging after amplification of PCR products with NdeI cleavage sites based on the dCAPS markers developed by ZmAPC4 and digested, agarose gel electrophoresis.
Detailed Description
The following detailed description of specific embodiments of the invention is, but it should be understood that the invention is not limited to specific embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention. The experimental methods described in the examples of the present invention are conventional methods unless otherwise specified, and materials, reagents, etc. used in the examples described below are commercially available.
Example 1
This example is an identification process of the starch content controlling gene ZmAPC4 in corn kernel.
1. Experimental materials
1. 261 maize inbred lines with abundant genetic diversity were provided by the professor Yan Jianbing to the university of agriculture in China.
2. Experimental method
1. Positioning group selection: the related population formed by 261 corn inbred lines with abundant genetic diversity is used as the positioning population of the starch content regulating genes in corn kernels.
2. Phenotype data acquisition:
(1) 261 corn inbred lines with abundant genetic diversity are continuously planted in Ledong county of Hainan province in winter three years in 2011-2013, and three biological repetitions are arranged in the field every year.
(2) After the corn is fully mature, the kernels are harvested and all 261 parts of kernels of the corn inbred line within three years 2011-2013 are used for starch content determination.
(3) The starch content in the corn kernel was measured using a near infrared spectrometer (model: MATRIX-1, manufactured by Brookfield, germany) and the procedure was as follows: each inbred line is selected, 30 corn kernel samples which are uniform and consistent are placed in a rotary sample cup with the diameter of 50mm, scanning is carried out on a near infrared spectrometer, in order to eliminate the influence of factors such as the granularity and uniformity of the samples on the spectrum, each inbred line is uniformly shaken after being measured for 1 time, the measurement is repeated for 3 times, and the average spectrum value is taken as the final measurement result.
(4) The BLUP value was calculated using lem4 package in R language software in combination with phenotype data of the starch content in 261 parts of inbred grain in three years 2011-2013, and the BLUP value of the starch content in 261 parts of inbred grain was used for subsequent Genome-wide association analysis (Genome-Wide Association Study, GWAS), and the GO (Gene Ontology) enrichment analysis result of the expression genes in corn grain screened using Genome-wide association analysis was shown in fig. 2.
(5) GWAS analysis was performed using BLUP values for starch content in 261 parts of maize inbred grain and Q-Q and manhattan plots were plotted (fig. 1). The related population used in the invention uses an Illumina MaizeSNP50 chip, simplified genome sequencing and an Affymetrix chip to sequence the whole genome of each maize inbred line in the earlier stage, and finally obtains about 55 ten thousand high-quality SNP markers (0.55M SNPs) with MAF more than or equal to 5% according to the genotyping result, and maps the SNP markers to a B73 RefGen_V2 reference genome. Due to the high linkage disequilibrium between certain markers, a moderate threshold value P.ltoreq.2.07×10 is used -5 As a basis for determining whether the SNP-phenotype is significantly associated.
GWAS was performed in tassel3.0 software using a General Linear Model (GLM) and introducing a Q model (control population structure) as an analytical model to obtain significant SNPs. LD analysis shows that the linkage disequilibrium attenuation distance of the set of markers is 50Kb, thus defining a total of 100Kb range of 50Kb upstream and downstream of the significant SNP as a significant site, and screening all genes in the significant site based on the B73 RefGen_V2 genome information. And obtaining obvious sites for regulating and controlling the starch content in corn kernels and possible candidate genes in the sites according to functional annotation of the genes, homologous gene information and enrichment analysis results.
The nucleotide sequence (CDS fragment) of the ZmAPC4 gene is shown as SEQ ID NO.1, and the encoded protein sequence is shown as SEQ ID NO. 2.
SEQ ID NO.1：
ATGGCGGAGGAGCAGATCGAAGAGGCGTCCATGGCCGCGGCGGCGGCCACGCCGTTCCAGCTCCAGTTCGACAAGCCCATCCCGTTCCAGATAAAAATGGCAGAATGGAATCCAGAGAAGGATCTGCTTGCTATGGTTACTGATGACTCCAAGGTTCTCCTCCATCGCTTCAATTGGCAAAGGCTTTGGACAATCTCTCCAGGGAAGTGCATCACATCAATTTGTTGGAGCCCTGATGGTAAAATAATAGCACTTGGCACAGAAGATGGTTTGATTCTTTTGCATGATGTGGAGAATGGGAAGATGCTAAGAACTACAAAATCCCACGATGTTGCCATTGTATCTTTGAATTGGGCAGAAGACGATCCACTGTCAAAGTCTGACAAGGATGAGTTTTTATCCTATGAAGACCGCACCACACGTTTCTTTCCTCCTCCTCCTGTGATGCCTAGGATAGGTGGACTACGTTCTGGAGATACTGGTCTTGCGGATGAGAACGAAGAAGCTATTCCAGAATTTTCTAGTGCTTCCTGTCAGCGTTTTAACATTCTGTGCAGTGGTGGCAAAGATGGTTGTGTTTGCTTCAGCGTTTTTGGAATATTTCCTGTTGGAAAGATAAACATAACTAAAATCCCAATCAATGTTGGATCCTCTAGAAAGAGTTACCAACTCCAGGATGCTTCAGTCAGCAAGGTCTCTTTATCAAGAAACCTCCAGAAATTTGTTATTCTGTGCTTTGGAAAGTTGGTTGATACCGACAACCTCTCTGACAGCTGCGAAAATTCTGGATTACATTGTCTCTACCTAGATACTTCTATCTTCTTCAACCGAAAAAATGAGCTGCATCAGGTGTCCCAACAAGCATCAAGTATTCAAGATATGGTTGAAGTTGTCCGTGCTTCTGTATCTTTAATATCCAAACAATGGTCAAACGCCATGAGTTTATTCCATGAGAAATTTAGTGCCTTGCCCAATCTGATATCTACACATGGAGTGGAATCTAGTTCTGAGGATGAGTTTCTTAGCCTTTTGTTTGGGACACGAACAAGTCCAGCCCTTCACCATTTTTTAGCTAGTTCACTTGGTGAAGCGGGTCTCAAGAGGATAGCCAAGGCTGTTGACAGTGCTGGAAGAGATATTCGTGGCATTATCACTGAGCATCTTCAGCCTGCAGTGGAGATTATTTCATTCAGACTCGCAGAATTAAGAGGCCTTTCAAGATGGCGTTCACGATTTCAAACTATCGGGTTAGATGGAAACCTTATTGATGGTGTAACTGAGAGTATAGGGATGCTAGTTGTTCAAGTGGAGCGCTTTTCAAGGGTGGCAGCCACTGTTGTTTATCTGTTTCAAAATTTCTTTGCCTGGGTTTTAAAGTCTGTTAGAATATTATTAAATGAACCTACCGACCAAGTTCCAGCAGCAAACAGTGAACTTGTGGTGATTTTTCTTAAGTTTCTTCTTGATAAGGATCCAATCAAACAACTACTTGAAGCAGATGAGAGAATTGAGTGTGATATGGATACTGCAAGACATGTAGAACAGCTAGTAGTTTTTGGAGGATTTACAGACACCCAATTTTTGGAAAAGAGTTTGGTAAATCAATTCAATGAATTGGAAGATAGCTTGAAGGAGGCTTTCTTGATGCCATTTACTGCTATCTCTTCGCAGATACAATGTCAAGGATTGCTTCCTTTATATCCTGTTACATCATCAGCTACCTTGTCATCATCCTGCTCGCCAACATCAATATCATTTTATAAGGATGAAGATTCATCGCATGAGGAATCCTCCTATAGCTTGACTGACTATGTATGCTTGAAGATACCTGATGGATCATTAAATAAAGTAAATTGCATCGGTGTGATAAAGGGCTCTGGTAACTGTTGTACTACCCTCAGCATGATGTCACTTTCAGGTTTTCTGTTGCATATACCTGACGGATATGAGTGTGTTGACCTGTCACTTTATAAGGACAACCAGGTAGTTTTACTACTGAGTGAAACGTCTTGTTCTGATAGCCCTGGAAAATCTTGGATGGTGATGCTGCAAATAGAGAACTTTTCCTTTATGCCACTCTCAGGAACATTCCCTGCAAATATTTACAGTTTGCAAAAACTGGTGGCACTCGATCTGCAATTGGACACAGATTATGGGAAAGTTCGTAGCATACCTCACACCGTATCTACTCCATTTGCAGTTAGTGCATCGAGAGGAGTAGCCTGTGTCTTTTCATCCCGGAGGCATGCCTTGGTTTATATCCTTGATGAGGATGAGGAAGCGGAGGGGGATGAAACTTCCGATATGGAGTGA
SEQ ID NO.2：
MetAlaGluGluGlnIleGluGluAlaSerMetAlaAlaAlaAlaAlaThrProPheGln
LeuGlnPheAspLysProIleProPheGlnIleLysMetAlaGluTrpAsnProGluLys
AspLeuLeuAlaMetValThrAspAspSerLysValLeuLeuHisArgPheAsnTrpGln
ArgLeuTrpThrIleSerProGlyLysCysIleThrSerIleCysTrpSerProAspGly
LysIleIleAlaLeuGlyThrGluAspGlyLeuIleLeuLeuHisAspValGluAsnGly
LysMetLeuArgThrThrLysSerHisAspValAlaIleValSerLeuAsnTrpAlaGlu
AspAspProLeuSerLysSerAspLysAspGluPheLeuSerTyrGluAspArgThrThr
ArgPhePheProProProProValMetProArgIleGlyGlyLeuArgSerGlyAspThr
GlyLeuAlaAspGluAsnGluGluAlaIleProGluPheSerSerAlaSerCysGlnArg
PheAsnIleLeuCysSerGlyGlyLysAspGlyCysValCysPheSerValPheGlyIle
PheProValGlyLysIleAsnIleThrLysIleProIleAsnValGlySerSerArgLys
SerTyrGlnLeuGlnAspAlaSerValSerLysValSerLeuSerArgAsnLeuGlnLys
PheValIleLeuCysPheGlyLysLeuValAspThrAspAsnLeuSerAspSerCysGlu
AsnSerGlyLeuHisCysLeuTyrLeuAspThrSerIlePhePheAsnArgLysAsnGlu
LeuHisGlnValSerGlnGlnAlaSerSerIleGlnAspMetValGluValValArgAla
SerValSerLeuIleSerLysGlnTrpSerAsnAlaMetSerLeuPheHisGluLysPhe
SerAlaLeuProAsnLeuIleSerThrHisGlyValGluSerSerSerGluAspGluPhe
LeuSerLeuLeuPheGlyThrArgThrSerProAlaLeuHisHisPheLeuAlaSerSer
LeuGlyGluAlaGlyLeuLysArgIleAlaLysAlaValAspSerAlaGlyArgAspIle
ArgGlyIleIleThrGluHisLeuGlnProAlaValGluIleIleSerPheArgLeuAla
GluLeuArgGlyLeuSerArgTrpArgSerArgPheGlnThrIleGlyLeuAspGlyAsn
LeuIleAspGlyValThrGluSerIleGlyMetLeuValValGlnValGluArgPheSer
ArgValAlaAlaThrValValTyrLeuPheGlnAsnPhePheAlaTrpValLeuLysSer
ValArgIleLeuLeuAsnGluProThrAspGlnValProAlaAlaAsnSerGluLeuVal
ValIlePheLeuLysPheLeuLeuAspLysAspProIleLysGlnLeuLeuGluAlaAsp
GluArgIleGluCysAspMetAspThrAlaArgHisValGluGlnLeuValValPheGly
GlyPheThrAspThrGlnPheLeuGluLysSerLeuValAsnGlnPheAsnGluLeuGlu
AspSerLeuLysGluAlaPheLeuMetProPheThrAlaIleSerSerGlnIleGlnCys
GlnGlyLeuLeuProLeuTyrProValThrSerSerAlaThrLeuSerSerSerCysSer
ProThrSerIleSerPheTyrLysAspGluAspSerSerHisGluGluSerSerTyrSer
LeuThrAspTyrValCysLeuLysIleProAspGlySerLeuAsnLysValAsnCysIle
GlyValIleLysGlySerGlyAsnCysCysThrThrLeuSerMetMetSerLeuSerGly
PheLeuLeuHisIleProAspGlyTyrGluCysValAspLeuSerLeuTyrLysAspAsn
GlnValValLeuLeuLeuSerGluThrSerCysSerAspSerProGlyLysSerTrpMet
ValMetLeuGlnIleGluAsnPheSerPheMetProLeuSerGlyThrPheProAlaAsn
IleTyrSerLeuGlnLysLeuValAlaLeuAspLeuGlnLeuAspThrAspTyrGlyLys
ValArgSerIleProHisThrValSerThrProPheAlaValSerAlaSerArgGlyVal
AlaCysValPheSerSerArgArgHisAlaLeuValTyrIleLeuAspGluAspGluGlu
AlaGluGlyAspGluThrSerAspMetGlu
Example 2
The embodiment is the screening of excellent haplotype of starch content regulating gene ZmAPC4 in corn kernel and the development of molecular marker.
Screening of excellent haplotype of starch content regulating gene ZmAPC4 in corn kernel
Based on genotype information of 0.55M SNPs, carrying out haplotype difference analysis by 261 parts of corn inbred grain starch content data and combining genotype data of the inbred lines in a ZmAPC4 interval (figure 3), screening corn inbred lines with higher starch content in the grain (as shown in table 1), and screening out excellent haplotypes with higher starch content, specifically, firstly eliminating unknown genotypes, and then dividing haplotypes by 261 parts of inbred grain starch content as phenotype data and combining genotypes of the inbred lines in the ZmAPC4 interval, wherein when the inbred coefficient quantity of a certain haplotype is less than 10, the haplotype is removed, thus obtaining 4 haplotypes in total, including Hap1 with nucleotide sequence shown as SEQ ID NO.3, hap2 with nucleotide sequence shown as SEQ ID NO.4, hap3 with nucleotide sequence shown as SEQ ID NO.5, and Hap4 with nucleotide sequence shown as SEQ ID NO.6, and the optimal haplotype of Hap4 is obtained. The maize inbred line carrying the Hap4 haplotype (table 1) has higher starch content and can provide excellent germplasm resources for breeding new maize varieties with higher starch content. Haplotypes of these inbred lines (containing the Hap4 haplotype) are called excellent haplotypes.
The nucleotide sequence of the excellent haplotype of the starch content regulating gene ZmAPC4 in the corn kernels is shown as SEQ ID NO. 6.
SEQ ID NO.3
AGTAACATTTCAA
SEQ ID NO.4
AGTAACATCACAG
SEQ ID NO.5
AGTAATGCTTTAG
SEQ ID NO.6
AGTAACATTTCAG
TABLE 1 Excellent haplotype maize inbred names with higher starch content
Sequence number Inbred line name
1 CIMBL48
2 CIMBL92
3 CIMBL50
4 CIMBL49
5 JH96C
6 GEMS14
7 04K5672
8 JI846
9 B151
10 LV28
11 CML432
Example 3
The embodiment is the application of a starch content regulating gene ZmAPC4 in corn kernels.
In order to verify the application value of the starch content regulating gene ZmAPC4 in corn kernels in actual production, a haplotype analysis of a single SNP was first performed according to the significant SNP chr4.s_175584318 (p=1.52E-05) highly associated with ZmAPC4 in GWAS analysis results, and as a result, as shown in fig. 4, there was a significant difference in starch content between corn inbred lines carrying two different alleles (TT or GG). Based on this significant SNP, molecular markers were developed for distinguishing starch content in corn inbred grain. In the specific embodiment, 8 corn inbred lines carrying GG genotype and having highest starch content and 8 corn inbred lines carrying TT genotype and having lowest starch content are selected based on 261 related populations, and the target fragment of 504bp containing significant SNP is amplified on DNA of leaves of the 16 corn inbred lines (table 2) by using primer pairs of SEQ ID NO.7 and SEQ ID NO. 8. dCAPS tags containing NdeI cleavage recognition sites were then designed using dCAPS Finder 2.0 software (http:// helix. Wust. Edu/dCAPS /) as shown in SEQ ID No.9 and SEQ ID No. 10. A251 bp fragment of interest containing NdeI cleavage recognition sites was then amplified from the 504bp fragment using the primer pair of SEQ ID NO.9 and SEQ ID NO. 10. The NdeI restriction endonuclease is used for cutting a 251bp target fragment, 4% agarose gel electrophoresis is used for identifying the product after the digestion, the fragment size of the product is observed in an ultraviolet gel imaging system, and corn inbred lines with different starch contents are distinguished according to the band type.
TABLE 2 16 maize inbred names and starch content for verification of molecular markers
Inbred line name Starch content in grain (%)
TIE7922 60.60
ZZ03 61.33
FCD0602 61.54
U8112 61.66
CML426 62.48
GEMS30 62.75
GEMS31 62.82
CIMBL127 63.33
SHEN135 69.42
CIMBL92 69.75
JY01 69.78
CIMBL48 70.46
CIMBL49 70.50
GEMS48 70.70
YE478 70.89
LV28 71.33
SEQ ID NO.7 (upstream primer): GTGGTAACAGCCTAACAGGTCACC
SEQ ID NO.8 (downstream primer): GCATTGTAACCAAGAAAGGTAGGCC
SEQ ID NO.9 (upstream primer): GCTATCGCATTTCTAATCGTGCATA
SEQ ID NO.10 (downstream primer): GCATTGTAACCAAGAAAGGTAGGCC
As shown in FIG. 5, after NdeI enzyme digestion, 8 corn selfing lacing patterns with lower starch content are obviously different from 8 corn selfing lacing patterns with higher starch content, which indicates that the dCAPS markers developed by the patent can be used for identifying and distinguishing the starch content in different corn selfing lines, and the line selection efficiency can be improved in the corn breeding process, namely, the starch content is determined by using the detection system of the invention through genotype identification without measuring the starch content after harvesting seeds, and the cultivation process of a new variety of high-quality corn with high starch content can be greatly accelerated through the molecular markers developed by the patent.
In conclusion, the nucleotide sequence (CDS fragment) of the ZmAPC4 gene obtained by screening is shown as SEQ ID NO.1, and the encoded protein sequence is shown as SEQ ID NO. 2. A batch of excellent corn germplasm with high starch content is screened through haplotype analysis of ZmAPC4, and in addition, the development of molecular markers can accelerate the corn breeding efficiency, so that the method has important significance in the actual production process.
While preferred embodiments of the present invention have been described, additional variations and modifications in those embodiments may occur to those skilled in the art once they learn of the basic inventive concepts. It is therefore intended that the following claims be interpreted as including the preferred embodiments and all such alterations and modifications as fall within the scope of the invention.
It will be apparent to those skilled in the art that various modifications and variations can be made to the present invention without departing from the spirit or scope of the invention. Thus, it is intended that the present invention also include such modifications and alterations insofar as they come within the scope of the appended claims or the equivalents thereof.

Claims (7)

1. Gene for regulating starch content in corn kernelZmAPC4Characterized in that the geneZmAPC4The nucleotide sequence of (2) is shown as SEQ ID NO. 1.
2. Gene for regulating starch content in corn kernelZmAPC4An expression product, characterized in that the geneZmAPC4Expression products, i.e.the genesZmAPC4The amino acid sequence of the coded protein is shown as SEQ ID NO. 2.
3. Gene for regulating starch content in corn kernelZmAPC4 SNP markers, characterized in that they are used for amplification comprisingZmAPC4Upstream primer sequence of primer pair of intramolecular specific SNP markerAs shown in SEQ ID NO.7, the sequence of the downstream primer is shown in SEQ ID NO. 8.
4. Gene for regulating starch content in corn kernelZmAPC4The haplotype is characterized by comprising 4 types of haplotypes, the nucleotide sequences of the haplotypes are shown as SEQ ID NO.3, SEQ ID NO.4, SEQ ID NO.5 and SEQ ID NO.6, and the SEQ ID NO.6 is an excellent haplotype.
5. The molecular marker for distinguishing the starch content in corn grains is characterized in that the upstream primer sequence of a primer pair of the molecular marker is shown as SEQ ID NO.9, and the downstream primer sequence is shown as SEQ ID NO. 10.
6. A gene for regulating and controlling starch content in corn kernel according to claim 1ZmAPC4The application in maize genetic breeding.
7. A gene for regulating and controlling starch content in corn kernel as claimed in claim 4ZmAPC4The application of the excellent haplotype in corn genetic breeding.
CN202310642357.9A 2023-06-01 2023-06-01 Gene ZmAPC4 for regulating starch content in corn kernel, expression product, SNP marker, excellent haplotype and application Pending CN116621961A (en)

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