CN111961739A - Specific primer group for detecting corn Dek41 gene and application thereof - Google Patents
Specific primer group for detecting corn Dek41 gene and application thereof Download PDFInfo
- Publication number
- CN111961739A CN111961739A CN202010534319.8A CN202010534319A CN111961739A CN 111961739 A CN111961739 A CN 111961739A CN 202010534319 A CN202010534319 A CN 202010534319A CN 111961739 A CN111961739 A CN 111961739A
- Authority
- CN
- China
- Prior art keywords
- dek41
- gene
- corn
- detecting
- specific primer
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 240000008042 Zea mays Species 0.000 title claims abstract description 53
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 52
- 235000002017 Zea mays subsp mays Nutrition 0.000 title claims abstract description 50
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 title claims abstract description 39
- 235000005822 corn Nutrition 0.000 title claims abstract description 38
- 238000009395 breeding Methods 0.000 claims abstract description 24
- 230000001488 breeding effect Effects 0.000 claims abstract description 24
- 239000012634 fragment Substances 0.000 claims abstract description 8
- 235000016383 Zea mays subsp huehuetenangensis Nutrition 0.000 claims description 13
- 235000009973 maize Nutrition 0.000 claims description 12
- 239000003147 molecular marker Substances 0.000 abstract description 18
- 241000196324 Embryophyta Species 0.000 abstract description 7
- 230000007547 defect Effects 0.000 abstract description 4
- 238000001514 detection method Methods 0.000 abstract description 4
- 238000009402 cross-breeding Methods 0.000 abstract description 3
- 230000009418 agronomic effect Effects 0.000 abstract description 2
- 108020004414 DNA Proteins 0.000 description 23
- 235000013339 cereals Nutrition 0.000 description 11
- 239000003550 marker Substances 0.000 description 10
- 238000000034 method Methods 0.000 description 8
- 235000018102 proteins Nutrition 0.000 description 8
- 102000004169 proteins and genes Human genes 0.000 description 8
- 238000005516 engineering process Methods 0.000 description 6
- 238000011161 development Methods 0.000 description 5
- 230000018109 developmental process Effects 0.000 description 5
- 235000013305 food Nutrition 0.000 description 5
- 239000000463 material Substances 0.000 description 5
- 102000053602 DNA Human genes 0.000 description 4
- 108020004682 Single-Stranded DNA Proteins 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 4
- 210000000349 chromosome Anatomy 0.000 description 4
- 210000001161 mammalian embryo Anatomy 0.000 description 4
- 102220533703 Histone-lysine N-methyltransferase SUV39H1_W64A_mutation Human genes 0.000 description 3
- 235000007244 Zea mays Nutrition 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 230000021759 endosperm development Effects 0.000 description 3
- 230000002068 genetic effect Effects 0.000 description 3
- 244000144972 livestock Species 0.000 description 3
- 235000016709 nutrition Nutrition 0.000 description 3
- 239000012188 paraffin wax Substances 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 210000005069 ears Anatomy 0.000 description 2
- 238000009396 hybridization Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000005204 segregation Methods 0.000 description 2
- 238000009331 sowing Methods 0.000 description 2
- 238000006467 substitution reaction Methods 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 108700028369 Alleles Proteins 0.000 description 1
- 241001483215 Balsas Species 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- 102000014171 Milk Proteins Human genes 0.000 description 1
- 108010011756 Milk Proteins Proteins 0.000 description 1
- 108700005079 Recessive Genes Proteins 0.000 description 1
- 102000052708 Recessive Genes Human genes 0.000 description 1
- 235000007241 Zea diploperennis Nutrition 0.000 description 1
- 235000017556 Zea mays subsp parviglumis Nutrition 0.000 description 1
- 241000172407 Zea mays subsp. huehuetenangensis Species 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000001133 acceleration Effects 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 238000010170 biological method Methods 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 244000240162 corn Species 0.000 description 1
- 230000002380 cytological effect Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- 235000012631 food intake Nutrition 0.000 description 1
- 238000012215 gene cloning Methods 0.000 description 1
- 238000003205 genotyping method Methods 0.000 description 1
- 239000012770 industrial material Substances 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 231100000518 lethal Toxicity 0.000 description 1
- 230000001665 lethal effect Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000013507 mapping Methods 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- 235000021239 milk protein Nutrition 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 230000010152 pollination Effects 0.000 description 1
- 102000054765 polymorphisms of proteins Human genes 0.000 description 1
- 244000144977 poultry Species 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000010153 self-pollination Effects 0.000 description 1
- 238000002864 sequence alignment Methods 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
- C12Q1/6895—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for plants, fungi or algae
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/13—Plant traits
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/156—Polymorphic or mutational markers
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Immunology (AREA)
- Mycology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Botany (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a specific primer group for detecting a corn Dek41 gene and application thereof. The pair of specific primers Dek41S1 of the left DNA fragment of Dek41 gene in the specific primer set amplifies the base sequence, and the pair of specific primers Dek41S11 of the right DNA fragment of Dek41 gene amplifies the base sequence. The two pairs of specific primer sets can be used for carrying out genotype identification on DNA of any tissue and any period of corn, and detecting the existence of Dek41 gene and the heterozygous or homozygous state in each individual plant of the filial generation of crossbreeding. The detection method has high accuracy and simple operation, and provides an important technical means for the DNA molecular marker-assisted breeding by using the Dek41 gene. The invention obtains and identifies the dominant molecular marker, which is the basis for carrying out DNA molecular marker assisted breeding on Dek41 gene for controlling the agronomic character of corn kernel defect.
Description
Technical Field
The invention relates to a specific primer group for detecting corn mutant genes and application thereof, in particular to a specific primer group for detecting corn Dek41 genes and application thereof.
Technical Field
Corn (zea mays) is a main food crop and feed crop in China in the world, and is known by the academic name zea mays L and English names maize or corn. The Chinese characters are more in name, which are called maize, corn and the like in old times, and are called corn in common now. Corn (Zea mays l.ssp. mays) is the most widely cultivated crop worldwide, a wild kindred species of the locality by the native inhabitants of the balsas river basin in the south of mexico more than 9000 years ago: maize-like/teosinte (Zea mays ssp. Parviglumis) is domesticated and selected. Calculated according to the yield and the sowing area, the corn is the first major food crop in China. Corn yield in china increased from 0.6 million tons in 1980 to 2.6 million tons in 2019 (national bureau of statistics, 2020). As the first large grain crop in China, the increase of the corn yield and the sowing area becomes an important factor for ensuring the national grain safety strategy and realizing the plan that the national grain production capacity in 2020 exceeds 6000 hundred million kilograms. Corn is an important feed and industrial material. Under the background of rapid development of Chinese economy and continuous acceleration of urbanization process, China has entered a stage of rapid increase of corn demand. First, the feed demand of corn is rapidly increasing. Along with the departure of the per-capita income level of Chinese residents, the per-capita food consumption structure of the residents is changing the demand of livestock products from carbohydrate food such as low-energy grains and vegetables to protein food such as high-energy livestock products, and the like, so that the demand of Chinese feed grains is continuously expanded and is in a 'rigid' growth situation. Therefore, the method puts higher demands on the improvement and innovation of the yield and the quality of the corn.
In order to improve the quality and the nutritional value of the corn, more and more scientific researchers at home and abroad begin to modify the quality of the corn by using a biological method from the beginning of the last century. High quality protein corn is produced under such conditions, and is a nutritionally balanced dual-purpose corn for food and feed. Compared with common corn, the content of lysine in the young protein corn is greatly increased, the nutritive value of the young protein corn is equivalent to milk protein, and the young protein corn can be used as livestock and poultry feed to improve the absorption and conversion of the protein and improve the yield. Meanwhile, the utilization rate of the animal to the feed protein can be improved, and the discharge of nitrogen in animal excrement is reduced, so that the pollution of the breeding industry to the environment is relieved. The principle of obtaining high-quality protein corn is that a mutant of partial grains in corn is used as a material, and the mutant is modified by means of a genetic research method, so that the nutritional value of the corn is improved. There are two major types of phenotypes studied in maize kernel mutants, Opaque and Defective kernel (Dek), respectively. The Opaque kernel phenotype is that the floury endosperm transparency is reduced, and the floury Opaque area is increased, so that the kernel hardness is reduced. Dek mutants are typically characterized by defects in endosperm development. Dek mutants often produce abnormal development of the embryo and endosperm and severe deficiencies in nutrient accumulation. During the whole kernel development period, Dek kernels are usually smaller than wild kernels, the endosperm development degree is slow, the filling degree is low, the seed coat is separated from the endosperm, the color of the seed coat is light gray, the surface of mature Dek kernels is shriveled and rough, the endosperm is often floury, the hardness is small, and the toughness is poor.
Dek41 is a recessive lethal Dek mutant. From the kernel phenotype, it was observed that the mutant kernel had reduced endosperm filling compared to the wild-type kernel, and the mutant kernel embryo was significantly smaller. The kernel mutants were severely shrunken during the maturation period, with significantly less kernel filler, as shown in figure 1. Although there is endosperm, it can be seen from the paraffin section material of the immature grain 15 days and 18 days after Dek41 pollination that the mutant embryo and endosperm are dysplastic relative to the wild type, there is a clear gap between endosperm and seed coat of the mutant grain, the embryo is obviously smaller, and the development is retarded, as shown in fig. 2. The analysis of the seeds in the aspects of phenotype, cytological section, seed nutritional ingredients and the like indicates that Dek41 is a new mutant; and allele detection analysis showed Dek41 to be caused by a monogenic recessive genetic mutation. The study of Dek41 mutant material can deepen our understanding of the cause of Dek mutant formation.
Dek41 mutant has not been well studied and utilized by cross breeding, because Dek41 recessive homozygous mutant has defect of kernel endosperm development, small endosperm and no growth, and most importantly, the cross breeding requires a long time. DNA molecular marker assisted breeding is a new breeding technology, and the technology is a modern breeding technology for indirectly selecting genes controlling target characters by using DNA molecular markers closely linked with the genes of the target characters. The technology can accurately and stably select target genes in early stage, and can overcome the problem of difficult identification by reusing recessive genes, thereby accelerating the breeding process and improving the breeding efficiency. Compared with conventional breeding, the technology can improve the breeding efficiency by 2-3 times.
The key of the DNA molecular marker assisted breeding technology is as follows:
(1) the obtained molecular marker should be a codominant primer group capable of distinguishing hybrid parents (homozygous Dek41 type and homozygous wild type) and hybrid offspring (F1 generation) thereof in the breeding process;
(2) the obtained co-dominant molecular marker (namely a primer group) and the control target character gene are linked, the closer the linkage is, the lower the probability of wrong selection in the offspring is, and if the marker is derived from the control target character gene, the molecular marker is used, the probability of wrong selection in the offspring is zero;
(3) the marker can distinguish the mutant from all the main breeding parents in China. In the past, no co-dominant DNA molecular marker (i.e., primer set) which is located on Dek41 gene and is completely linked with it was obtained, so that it was impossible to perform DNA molecular marker-assisted selection on the target trait controlled by the gene, and in the past, it was also possible to obtain an analytical marker which can distinguish the mutant from all the major breeding parents in China. Therefore, the acquisition and identification of the marker are the basis for carrying out DNA molecular marker assisted breeding on Dek41 genes for controlling the agronomic traits of corn kernel defects (Dek).
Disclosure of Invention
One of the purposes of the invention is to provide a specific primer group for detecting the corn Dek41 gene.
The invention also aims to provide application of the specific primer group in detecting and distinguishing hybrid parents in a corn breeding process.
The invention also aims to provide application of the specific primer group in detecting and distinguishing wild type inbred lines B73, Zheng 58, Mo17, Chang 7-2, PH4CV and PH6WC types and hybrid offspring types thereof.
In order to achieve the purpose, the invention adopts the following technical scheme:
a specific primer group for detecting a corn Dek41 gene, wherein a pair of specific primers Dek41S1 for amplifying a DNA fragment on the left side of the Dek41 gene in the specific primer group have the following base sequences:
the forward primer from 5 'end to 3' end is: GACGCCACTGTTCTCTCATGC, respectively;
the reverse primer is from 5 'end to 3' end: TGCCAGCTATGTCCTGAACTTC, respectively;
the base sequences of a pair of specific primers Dek41S11 for amplifying the right DNA fragment of the Dek41 gene are as follows:
the forward primer from 5 'end to 3' end is: CACCTGCCTAAGCTGCCTTG, respectively;
the reverse primer is from 5 'end to 3' end: GCCTCTCTATCGTGGTTTGCTG are provided.
Use of the specific primer set for detecting the maize Dek41 gene according to claim 1 in detecting and distinguishing the crossed parents and their hybrid progeny types in maize breeding.
Use of the specific primer set for detecting maize Dek41 gene according to claim 1 for detecting and differentiating wild type inbred lines B73, Zheng 58, Mo17, Chang 7-2, PH4CV and PH6WC type and their hybrid progeny type.
The invention provides 2 codominant DNA specific primer groups which are positioned at two sides of the physical position of Dek41 gene chromosome and are closely linked with the same, and a method for carrying out molecular marker-assisted selection on Dek41 gene by utilizing the two codominant DNA specific primer groups. The two specific primer sets can be used for carrying out genotype identification on DNA of corn at any period and in any tissue, and detecting the existence of Dek41 gene and the heterozygous or homozygous state in each individual plant of hybrid breeding filial generation. The detection method has high accuracy and simple operation, and provides an important technical means for the DNA molecular marker-assisted breeding by using the Dek41 gene.
Drawings
FIG. 1 is a graph of representative ears from hybrid F2, and a comparison of mutant and wild type kernels. Wherein panel a is a diagram of Dek41 hybrid F2 representative isolated ears, and panel B is a comparison of wild type grain to Dek41 pure and mutant grain.
Fig. 2 is a picture of paraffin section materials of wild type and mutant kernel. Wherein, the picture A and the picture B are respectively the paraffin section material pictures of the wild type and the mutant grain 15 days and 18 days after the self-pollination of the Dek41 hybrid F1 generation.
FIG. 3 is a schematic diagram of the F2 population construction for Dek41 gene cloning.
FIG. 4 is a graph of the segregation of marker Dek41S1 of the present invention in the W64A background F2 population. Dek41/Dek41 in the figure are homozygous mutants; dek41/+ is a hybrid; +/+ is homozygous wild type.
FIG. 5 is a graph of the segregation of marker Dek41S11 of the present invention in the W64A background F2 population. Dek41/Dek41 in the figure are homozygous mutants; dek41/+ is a hybrid; +/+ is homozygous wild type.
FIG. 6 is a schematic diagram showing the positions of molecular markers on chromosome 7 of maize in accordance with the present invention.
FIG. 7 shows the polymorphism of marker Dek41S1 among different inbred lines.
FIG. 8 shows the polymorphism of marker Dek41S11 among different inbred lines.
Detailed Description
The invention is further illustrated below with reference to specific examples. The following examples are intended to illustrate the invention only and are not intended to limit the scope of the invention. The experimental procedures for the specific experimental conditions not specified in the examples below are generally carried out according to conventional conditions, such as those described in Molecular Cloning (A Laboratory Manual,3rd ed.) or Plant Molecular Biology-Laboratory Manual (Plant Molecular Biology-A Laboratory Manual, eds., memory S.Clark, Springer-verlag Berlin Heidelberg,1997), or according to the conditions recommended by the manufacturer.
The first embodiment is as follows:
acquisition of two co-dominant molecular markers:
this example demonstrates the location of the Dek41 gene on chromosome 7 in maize by classical genetic mapping. Genomic DNA of individual plants of homozygous mutant, wild type and F2 populations was extracted by constructing the F2 population for plants of heterozygous mutant and homozygous wild type near position Dek41, see fig. 3. Primers were designed based on the known genomic sequence of B73 (http:// www.genome.arizona.edu/fpc/mail). Through sequencing and sequence alignment, the sequence difference between the homozygous mutant and the homozygous wild-type parent is obtained, and through the development of markers of the sequenced sequence, the co-dominant molecular markers Dek41S1 and Dek41S11 capable of distinguishing the hybridized parents and the hybridized filial generations (F1 generation) in the breeding process are obtained. In this example, the hybrid parents are homozygous Dek41 type and homozygous wild type, and the filial generation of the hybrid parent is F1 generation.
In this example, in the specific primer set for detecting the maize Dek41 gene, the base sequence of the pair of specific primers Dek41S1 for amplifying the DNA fragment on the left side of the Dek41 gene is:
the forward primer from 5 'end to 3' end is: GACGCCACTGTTCTCTCATGC, respectively;
the reverse primer is from 5 'end to 3' end: TGCCAGCTATGTCCTGAACTTC, respectively;
the base sequences of a pair of specific primers Dek41S11 for amplifying the right DNA fragment of the Dek41 gene are as follows:
the forward primer from 5 'end to 3' end is: CACCTGCCTAAGCTGCCTTG, respectively;
the reverse primer is from 5 'end to 3' end: GCCTCTCTATCGTGGTTTGCTG are provided.
Example two:
determination of linkage relationship between molecular markers Dek41S1, Dek41S11 and Dek41 respectively:
the DNA of seeds in an F2 population obtained after Dek41/+ background hybridization with W64A background hybridization is respectively extracted and used for analyzing the linkage relation between molecular markers Dek41S1 and Dek41S11 and Dek41H respectively, the genotypes of wild phenotypes comprise +/+ and Dek41/+, and the genotype of a Dek41 homozygous mutant is Dek41/Dek 41. The samples were used to analyze the genetic linkage of molecular markers Dek41S1, Dek41S11 to Dek41 gene, respectively, see FIGS. 4 and 5. The analysis result shows thatTwo markers were linked to the Dek41 gene, and analysis of the large population showed that the crossover of marker Dek41S1 was 1.0X 10 in the segregating population on a W22 background-3The exchange rate of marker Dek41S11 was 1.2X 10-2. The molecular markers Dek41S1, Dek41S11 are physically located on the chromosome relative to Dek41 gene, respectively, as shown in FIG. 6.
Example three:
polymorphism identification of molecular markers among different parents:
genomic DNAs of types B73, Zheng 58, Mo17, Chang 7-2, PH4CV and PH6WC of inbred lines widely used in Dek41 and 8 breeding were extracted, and polymorphisms among different varieties were analyzed by PCR reaction. The results show that the molecular marker Dek41S1 can distinguish Dek41 from B73 and Zheng 58 inbred lines, and the molecular marker Dek41S11 can distinguish Dek41 from B73, Chang 7-2, Mo17 and Zheng 58 inbred lines, as shown in FIG. 7 and FIG. 8. The results prove that the molecular marker Dek41S1 and the molecular marker Dek41S11 can accurately detect the presence of Dek41 genes in B73 and Zheng 58 background populations, and the misselection rate in auxiliary selection is 1.2 multiplied by 10-5And the selection requirement can be met.
In conclusion, the two pairs of specific primer sets of the above embodiments can be used for genotyping DNA of corn at any time and in any tissue, and detecting the presence or absence and heterozygous or homozygous state of Dek41 gene in each individual plant of hybrid breeding progeny. The detection method has high accuracy and simple operation, and provides an important technical means for the DNA molecular marker-assisted breeding by using the Dek41 gene.
The embodiments of the present invention have been described with reference to the accompanying drawings, but the present invention is not limited to the embodiments, and various changes and modifications can be made according to the purpose of the invention, and any changes, modifications, substitutions, combinations or simplifications made according to the spirit and principle of the technical solution of the present invention shall be equivalent substitutions, as long as the purpose of the present invention is met, and the present invention shall fall within the protection scope of the present invention without departing from the technical principle and inventive concept of the present invention.
Sequence listing
<110> university at Shanghai
<120> specific primer group for detecting corn Dek41 gene and application thereof
<160> 4
<170> SIPOSequenceListing 1.0
<210> 1
<211> 22
<212> Single-stranded DNA
<213> Artificial sequence ()
<400> 1
<210> 2
<211> 22
<212> Single-stranded DNA
<213> Artificial sequence ()
<400> 2
<210> 3
<211> 20
<212> Single-stranded DNA
<213> Artificial sequence ()
<400> 3
<210> 4
<211> 20
<212> Single-stranded DNA
<213> Artificial sequence ()
<400> 4
Claims (3)
1. A specific primer group for detecting a corn Dek41 gene, which is characterized in that: the base sequences of a pair of specific primers Dek41S1 for amplifying the DNA fragment on the left side of the Dek41 gene in the specific primer group are as follows:
the forward primer from 5 'end to 3' end is: GACGCCACTGTTCTCTCATGC, respectively;
the reverse primer is from 5 'end to 3' end: TGCCAGCTATGTCCTGAACTTC, respectively;
the base sequences of a pair of specific primers Dek41S11 for amplifying the right DNA fragment of the Dek41 gene are as follows:
the forward primer from 5 'end to 3' end is: CACCTGCCTAAGCTGCCTTG, respectively;
the reverse primer is from 5 'end to 3' end: GCCTCTCTATCGTGGTTTGCTG are provided.
2. Use of the specific primer set for detecting the gene Dek41 in maize according to claim 1 for detecting and distinguishing the hybrid parents and their hybrid progeny types in maize breeding.
3. Use of the specific primer set for detecting the gene of maize Dek41 according to claim 1 for detecting and differentiating the types of wild-type inbred lines B73, Zheng 58, Mo17, Chang 7-2, PH4CV and PH6WC and their hybrid progeny types.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010534319.8A CN111961739B (en) | 2020-06-12 | 2020-06-12 | Specific primer group for detecting Dek gene of corn and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010534319.8A CN111961739B (en) | 2020-06-12 | 2020-06-12 | Specific primer group for detecting Dek gene of corn and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111961739A true CN111961739A (en) | 2020-11-20 |
| CN111961739B CN111961739B (en) | 2024-03-19 |
Family
ID=73360392
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010534319.8A Active CN111961739B (en) | 2020-06-12 | 2020-06-12 | Specific primer group for detecting Dek gene of corn and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111961739B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114292947A (en) * | 2021-12-31 | 2022-04-08 | 中国农业科学院作物科学研究所 | CAPS mark for identifying number of layers of corn gruel layer and detection method thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108034743A (en) * | 2017-12-06 | 2018-05-15 | 上海大学 | For detecting specific primer and its application of corn Dek6 genes |
| CN108411025A (en) * | 2018-04-28 | 2018-08-17 | 上海大学 | The specific primer group of augmentation detection corn dek33 gene molecular labelings and its application |
| CN108660242A (en) * | 2018-04-28 | 2018-10-16 | 上海大学 | The specific primer group of augmentation detection corn dek10 gene molecular labelings and its application |
-
2020
- 2020-06-12 CN CN202010534319.8A patent/CN111961739B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108034743A (en) * | 2017-12-06 | 2018-05-15 | 上海大学 | For detecting specific primer and its application of corn Dek6 genes |
| CN108411025A (en) * | 2018-04-28 | 2018-08-17 | 上海大学 | The specific primer group of augmentation detection corn dek33 gene molecular labelings and its application |
| CN108660242A (en) * | 2018-04-28 | 2018-10-16 | 上海大学 | The specific primer group of augmentation detection corn dek10 gene molecular labelings and its application |
Non-Patent Citations (3)
| Title |
|---|
| CHENGUANG ZHU等: "Maize pentatricopeptide repeat protein DEK41 affects cis-splicing of mitochondrial nad4 intron 3 and is required for normal seed development" * |
| RU CHANG REN;LI LI WANG;LIN ZHANG;YA JIE ZHAO;JIA WEN WU;YI MING WEI;XIAN SHENG ZHANG;XIANG YU ZHAO;: "DEK43 is a P-type pentatricopeptide repeat(PPR)protein responsible for the Cis-splicing of nad4 in maize mitochondria" * |
| 石慧敏;蒋成功;王红武;马庆;李坤;刘志芳;吴宇锦;李树强;胡小娇;黄长玲;: "玉米籽粒突变体dek48的表型鉴定与基因定位" * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114292947A (en) * | 2021-12-31 | 2022-04-08 | 中国农业科学院作物科学研究所 | CAPS mark for identifying number of layers of corn gruel layer and detection method thereof |
| CN114292947B (en) * | 2021-12-31 | 2023-09-29 | 中国农业科学院作物科学研究所 | CAPS mark for identifying number of corn aleurone layers and detection method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111961739B (en) | 2024-03-19 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US6300541B1 (en) | Soybean sudden death syndrome resistant soybeans, soybean cyst nematode resistant soybeans and methods of breeding and identifying resistant plants | |
| CN109735652B (en) | Wheat stripe rust resistant gene QYr.nwafu-6BL.2 linked KASP molecular marker, primer and application | |
| EP3561079B1 (en) | Soybean anti-pod-shattering major qtlqpd08-1, and mapping method and application thereof | |
| CN116497129B (en) | Gene molecular marker related to stichopus japonicus growth traits and application thereof | |
| CN110747288A (en) | Rice large grain gene function marker and application | |
| CN107400706B (en) | Specific primer group of molecular marker for amplification detection of maize opaque11 gene and application thereof | |
| CN111893209B (en) | Indel site detection marker related to thousand grain weight of wheat and application thereof | |
| CN113736910B (en) | Linkage molecular marker of peanut single plant pod number major QTL locus qPN and application thereof | |
| CN109609687B (en) | KASP marker primer combination for detecting watermelon fusarium wilt resistance and application thereof | |
| CN109486829B (en) | Rice semi-dwarf gene sd1 allele and identification method thereof | |
| CN109111511B (en) | Cultivation method of super-long rice | |
| CN111961739B (en) | Specific primer group for detecting Dek gene of corn and application thereof | |
| CN103834647A (en) | SSR marker Xgwm537 in close linkage with wheat dwarf gene RhtDC20 and application thereof | |
| CN115232823B (en) | Cabbage mustard mushroom leaf development related gene and application thereof | |
| CN107287210B (en) | Rice appearance quality gene qAQ7 and molecular marking method and application thereof | |
| US10028459B2 (en) | Tomato plants with improved disease resistance | |
| CN108660242B (en) | Specific primer group of molecular marker for amplification detection of corn dek10 gene and application thereof | |
| CN106929590B (en) | Specific primer group for amplifying and detecting molecular marker of corn 5512J gene and application thereof | |
| Karakotov et al. | Modern issues of sugar beet (Beta vulgaris L.) hybrid breeding | |
| CN108411025B (en) | Specific primer group of molecular marker for amplification detection of corn dek33 gene and application thereof | |
| CN111996280B (en) | SNP marker co-separated from brassica napus dwarf compact trait and application thereof | |
| CN110283924B (en) | Specific primer group for detecting maize 3906H gene and application thereof | |
| CN108315463B (en) | Primer group for detecting corn 1768 gene and application thereof | |
| CN111004857A (en) | Molecular marker primer of major QTL (quantitative trait locus) site of soybean branch number and application of molecular marker primer | |
| CN117887885B (en) | Soybean oil content-related major single nucleotide polymorphism site and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |