CN116814839B - Molecular marker AhyCs1 closely linked with peanut seed coat color speckles and application thereof - Google Patents
Molecular marker AhyCs1 closely linked with peanut seed coat color speckles and application thereof Download PDFInfo
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- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
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Abstract
The invention discloses molecular marker AhyCs1 closely linked with peanut seed coat color speckles and application thereof, and belongs to the technical field of agricultural biology, wherein a specific primer pair of the molecular marker AhyCs1 comprises a forward primer and a reverse primer shown as SEQ ID No. 1-2. The molecular marker AhyCs1 can be used for identifying peanuts with color speckle seed coats, can be used for determining the seed coat color of next-generation harvesting materials in advance by detecting the DNA of seeds and leaves of the peanuts, improves the breeding efficiency, is beneficial to fine positioning, separation and cloning of genes, and has important application value for molecular breeding and quality improvement of the peanuts.
Description
Technical Field
The invention relates to the technical field of agricultural biology, in particular to a molecular marker AhyCs1 closely linked with peanut seed coat color speckles and application thereof.
Background
Peanuts are important oil crops and economic crops in China, the annual planting area of the peanuts in China is about 7000 ten thousand mu, and the yield is about 1700 ten thousand tons. The peanut is rich in fatty acid, protein, microelements such as calcium, iron, zinc, selenium and the like, vitamin E, functional active substances such as resveratrol and the like, and has important health care function. In China, peanuts are also called as ' long fruits ', and the efficacy of the peanuts is recorded in Ming Dynasty's herbal compendium ', xin Lin medical science ', lu Chuan Ben Cao's and national Chinese herbal medicine assembly '. In abroad, a great deal of researches also show that the peanuts have remarkable effects in preventing and treating cardiovascular diseases, treating malnutrition and the like. For example, a study in New England journal of medicine (The New England Journal ofMedicine) shows that eating nuts daily can reduce mortality from heart disease by 29% and even eating peanuts twice a week can reduce risk by 24% (Bao et al 2013). Peanut butter with a special formula mainly used for peanuts has become a main therapeutic instant nutritional food for treating children malnutrition in the united nations in africa. To date, the peanut butter program (Project Peanut Butter) has cured 100-thousand malnourished children in 2004.
Peanut seed coats, also known as peanut red coats, are important carriers of peanut nutrients. Peanut seed coats are rich in natural pigments, especially anthocyanin, which has various biological effects of inhibiting free radicals, resisting oxidation and the like (Zhang Yufang, 2003). In addition, the peanut seed coats are also called peanut red coats, so that the traditional Chinese medicine has the effect of stopping bleeding and treating hemophilia, and the peanut red coat pigment is a pure natural pigment, is nontoxic and safe, and is an antioxidant natural pigment which is difficult to obtain in the market. The color of the seed coats of the peanuts is mainly pink and red, and the peanut varieties with the color spot seed coats are less and the general yield is not high. The color depth of the peanut seed coats is closely related to anthocyanin accumulation, and the color spot areas of the peanut seed coats are enriched with more anthocyanin. Anthocyanin has various biological effects of resisting oxidation and inhibiting free radical accumulation. In recent years, market demand for color spot seed coat peanuts is increasing. However, the existing color spot varieties only have few varieties such as 'Yunnan colorful peanuts'. The peanut varieties with color spots do not exist in main peanut producing areas such as Henan and Shandong, and the yield of the existing peanut varieties with color spots is lower than that of the conventional varieties. Therefore, the cultivation of the high-yield colored spot seed coat peanuts is a fundamental way for meeting the market demand.
The seed coat of peanuts is also developed from beads, and the color of the seed coat is developed at alternate generations (Zhao et al 2020). In addition, peanuts belong to crops with 'flowering on the ground and fruiting on the ground', and the color of seed coats can be counted after harvesting. The characteristics greatly limit the cultivation of peanut varieties with color spot seed coats by the traditional breeding means. For the color spot seed coat character, the genotype of the cotyledon is cut off, so that the seed coat color (enemy calm and the like, 2018) of the next-generation harvested material can be determined in advance by detecting the genotype of the current-generation leaf, the seed coat color of the current-generation harvested material can be determined in advance by detecting the genotype of the current-generation leaf, the process of cultivating the new variety of the deep color spot peanut can be accelerated, and the breeding efficiency is improved. However, there are no reports on peanut color spot seed coat related genes and molecular markers.
Disclosure of Invention
The invention aims to provide molecular markers AhyCs1 closely linked with peanut seed coat color speckles and application thereof, so as to solve the problems in the prior art, the molecular markers can effectively identify the color of the peanut color speckles seed coats, can determine the seed coat color of a next-generation harvesting material in advance by detecting the DNA of seeds and leaves of the peanut, improve the breeding efficiency, are beneficial to fine positioning, separation and cloning of genes, and have important application values for molecular breeding and quality improvement of the peanut.
In order to achieve the above object, the present invention provides the following solutions:
the invention provides a molecular marker AhyCs1 closely linked with peanut seed coat color speckles, wherein a specific primer pair of the molecular marker AhyCs1 comprises:
a primer with a nucleotide sequence shown as SEQ ID No. 1;
the nucleotide sequence is shown as a primer shown as SEQ ID No. 2.
The invention also provides a method for identifying the color of peanut seed coats by using the molecular marker AhyCs1, which comprises the following steps:
extracting DNA of peanut leaves or seeds to be detected as a template, carrying out PCR amplification by using the specific primer pair, then carrying out gel electrophoresis detection on the amplified product, and judging the seed coat color of the peanut to be detected according to the gel electrophoresis result.
Further, if the gel electrophoresis result shows that the characteristic band with the size of 233bp appears, the peanut and the seeds of the offspring thereof to be detected are judged to be color spot streak skin.
Further, if the gel electrophoresis result only shows a characteristic band with the size of 233bp, judging that seeds of the peanut and the offspring thereof to be detected are obvious color spot streak skins; if the gel electrophoresis result shows that characteristic bands with the sizes of 229bp and 233bp are simultaneously generated, judging that seeds of the peanut and the offspring thereof to be detected are light colored spot striped skins; if the gel electrophoresis result only shows a characteristic band with the size of 229bp, judging that the seeds of the peanut and the offspring thereof to be detected are not the color spot streak skin.
Further, the PCR system for PCR amplification is as follows: 1. Mu.L of DNA template, 0.5. Mu.L of forward primer and reverse primer each, 0.5. Mu.L of dNTPMix, 2.0. Mu.L of 10 XTaq Buffer, mgCl 2 2.0. Mu.L of Taq enzyme, 0.20. Mu.L of water was added to 20. Mu.L.
Further, the PCR amplification PCR procedure is as follows: pre-denaturation at 95℃for 3min;94℃for 30s,56℃for 30s,72℃for 20s,35 cycles; extending at 72℃for 5min. .
Further, the gel electrophoresis detection comprises 8% non-denaturing polyacrylamide gel electrophoresis.
The invention also provides a kit for identifying peanut seed coat color speckles, which comprises a specific primer pair shown as SEQ ID No. 1-2.
The invention also provides application of the molecular marker AhyCs1, wherein the molecular marker AhyCs1 is used for peanut breeding and/or peanut quality improvement.
Further, the method is used for identifying the color zebra peanuts of the seed coats.
The invention discloses the following technical effects:
the molecular marker AhyCs1 closely linked with the peanut color speckle seed coats can be used for identifying the peanut with the color speckle seed coats, and the seed coat color of the next-generation harvesting material can be determined in advance by detecting the DNA of the seeds and leaves of the peanut, so that the breeding efficiency is improved. In addition, peanut is a heterotetraploid, alleles of the A and B subgenomic genes are highly homologous, and many markers are difficult to effectively distinguish the A and B subgenomic genes, and the molecular markers are screened through multiple scientific experiments and fumbling, so that the results are reliable and the reliability is high.
The molecular marker is a simple PCR marker, has simple technical requirements, can realize identification directly through PCR amplification and electrophoresis without enzyme digestion, purification, recovery and other steps compared with CAPS molecular markers, has lower requirements on instrument operation, can be operated by adopting a conventional instrument of a conventional experiment, and is more easily accepted by people.
The molecular marker can effectively identify the color of the color speckle seed coat of the peanut, is favorable for fine positioning, separation and cloning of genes on one hand, and has important application value for molecular breeding and quality improvement of the peanut on the other hand.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are needed in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 shows the initial mapping of genes controlling peanut color spot seed coat color by whole genome resequencing combined with cluster separation analysis; a: constructing an extreme pool by utilizing the segregating population for BSA-seq analysis; b: distribution of markers linked to colored speckle seed coats on peanut genomes;
FIG. 2 is a diagram showing the control of fine localization of peanut color spot seed coat genes and the development of closely linked molecular markers;
FIG. 3 shows the results of verification of molecular markers AhyCs1 in peanut materials with different colors;
FIG. 4 is a sample of peanut seeds and a germination test of the peanut seeds after the sample; a: complete seed control; b: cutting the cotyledon seed; c: germination of uncut cotyledons; d: cutting off germination of cotyledons;
FIG. 5 shows the authenticity identification of the hybrid F1 generation by using the molecular marker AhyCs 1; true filial offspring in box;
FIG. 6 is a breeding flow chart of molecular marker assisted recurrent selection and partial high-yield color spot seed coat peanut new strain obtained by molecular marker screening.
Detailed Description
Various exemplary embodiments of the invention will now be described in detail, which should not be considered as limiting the invention, but rather as more detailed descriptions of certain aspects, features and embodiments of the invention.
It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. In addition, for numerical ranges in this disclosure, it is understood that each intermediate value between the upper and lower limits of the ranges is also specifically disclosed. Every smaller range between any stated value or stated range, and any other stated value or intermediate value within the stated range, is also encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.
Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.
It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the invention described herein without departing from the scope or spirit of the invention. Other embodiments will be apparent to those skilled in the art from consideration of the specification of the present invention. The specification and examples of the present invention are exemplary only.
As used herein, the terms "comprising," "including," "having," "containing," and the like are intended to be inclusive and mean an inclusion, but not limited to.
Example 1 design of molecular marker AhyCs1 controlling the location of peanut colored speckle seed coat color Gene and close linkage with peanut colored speckle seed coat
In order to locate and control peanut color spot seed coat color genes, the inventor utilizes a pure (pink) seed coat peanut variety (female parent) G06G (from a Shandong province agricultural crop germplasm resource library) to hybridize with a peanut variety (male parent) seven-color peanut (QCHS, from a Shandong province agricultural crop germplasm resource library) with color spot seed coats, so as to construct a separation population, and genetic analysis shows that the color spots exist and are not controlled by a single gene.
By phenotyping F2 and F3 generation stain seed coats, 30 offspring lines with significant stains and 30 solid (pink) color were selected to construct extreme pools of stain seed coats and pink seed coats, respectively, and full genome resequencing was performed with the parent G06G and sevens peanut, sequencing amounts of 32Gb and 92Gb for the parent and extreme pools, respectively (a of fig. 1). The resequencing data were analyzed using cluster segregation analysis (BSA) and the distribution of candidate linked SNP sites on the peanut genome was counted, and a large number of SNP sites were found to be present on chromosome 6 (chr.06) (B of fig. 1), indicating that genes controlling the color of peanut colored-spot seed coats may be on chromosome 6.
Further developing a marker in a candidate interval according to the initial positioning result of the BSA-seq, detecting genotypes in all strains of the segregating population, and constructing a local high-density genetic linkage map of the candidate interval. Key genes controlling peanut color zebra seed coat color were located and locked within the 24745546-49876762bp interval of chromosome 6 (fig. 2) by linkage analysis and comparative genomics analysis in combination with phenotypic statistics for each strain seed coat color. Meanwhile, the inventor discovers that the colors of the marked AhyCs1 and the color speckle seed coats are closely linked, and the specific primers of the AhyCs1 are utilized to verify in different color germplasm materials (figure 3), and as can be seen from figure 3, the peanuts with the color speckle seed coats (dark and obvious color speckle seed coats) are 233bp characteristic bands, the peanuts with the intermediate seed coats (light color speckle seed coats) are 229 and 233bp characteristic bands, and the peanuts with the pink seed coats are 229bp characteristic bands. The proving mark AhyCs1 is an effective mark for judging the color of peanut seed coats.
Specific primer pairs for AhyCs1 include forward primers: 5'-GTATTGAAGAAAGCTCGGAATT-3' (SEQ ID No. 1) and reverse primer: 5'-AGCGGTGAAGAGGAAGATGA-3' (SEQ ID No. 2).
Example 2 quick cultivation of high yield seed-coat peanut New variety of colored spots Using molecular marker AhyCs1
The method for selecting and combining backcross breeding by using the molecular marker AhyCs1 can improve the common peanut (recurrent parent) into a new peanut variety with a color spot seed coat in about 3 years, and can realize the directional improvement of the seed coat color from pink to the color spot seed coat on the premise of keeping most of excellent characters of the original peanut variety by keeping the genetic background of more than 97% of recurrent parent.
The specific steps of color improvement of the peanut color spot seed coats by taking a common peanut seed and flower breeding 23 (HY 23, derived from a farm crop germplasm resource pool in Shandong province) as a recurrent parent and taking a color spot seed coat peanut variety seven-color peanut (QCHS) as a donor male parent include:
(1) Hybridization
The hybrid is carried out by taking HY23 as a female parent (recurrent parent) and taking a color spot seed coat peanut variety QCHS as a male parent. The hybridization method is as follows: castration begins a few days after parent HY23 flowers, typically 16 per day: emasculation was performed after 00. The basal part of the flower bud is pinched by the thumb and middle finger of the left hand, the calyx, the flag valve and the wing valve are gently pulled out by the forceps held by the right hand, the pulled-out petals are pressed by the index finger and thumb of the left hand, then the back bending part of the keel valve is gently pressed by the forceps to expose the flower pistil, the anther of 8 stamens is plucked out by the forceps once or a plurality of times, the column heads of the pistil are not damaged, and the keel valve is pushed back to the original position by the fingers. Emasculated flowers were artificially pollinated 5:00-9:00 a day 2 earlier. Flowers of male parent QCS are collected before pollination, pollen of the male parent flowers is extruded by forceps, when pollination is carried out, the flowers with male flowers removed are held by a left index finger and a middle finger, a right thumb or forceps are used for lightly extruding keel petals, pistil stigma is exposed, and the tips of the forceps are used for dipping pollen and coating the pollen on the stigma.
(2) Hybridization F 1 Authentication of authenticity
Hybridization F for harvesting by using molecular marker AhyCs1 1 And (5) authenticating the authenticity of the generation. The method comprises the following steps:
sampling: all pods of the female parent plants are harvested, and after airing, all seeds harvested are numbered. Then, a part of the seed coat was removed by a scalpel, and then a part of the cotyledon tissue (about 30 mg) was excised, placed in a 1.5mL centrifuge tube, and simultaneously placed in a magnetic bead. And (5) placing the rest peanut seeds into a refrigeration house for preservation, and planting the peanut seeds into a field after detection. Experiments prove that the germination rate of peanut seeds is not affected after partial tissues are excised (fig. 4).
DNA extraction: the specific method for extracting DNA of peanut seeds to be detected comprises the following steps:
(1) The 1.5mL centrifuge tube filled with peanut tissue is rapidly cooled by liquid nitrogen and then ground;
(2) Preheating CTAB extract (2% CTAB,1.4mol/LNaCl,20mmo/L EDTA (pH 8.0), 100 mmole/LTris-HCl (pH 8.0), 2% pvp-40) in a water bath at 65 ℃;
(3) Estimating the mass of a sample tissue, adding 700 mu L of preheated CTAB extract into every 200mg of sample, quickly and uniformly mixing, and carrying out warm bath for 10-30 min at 65 ℃ for 2-5 times;
(4) Adding phenol/chloroform/isoamyl alcohol (volume ratio is 12:12:1) with volume of 1 time, and uniformly mixing;
(5) Centrifuging at 12000rpm at room temperature for 10min;
(6) Transferring the supernatant to a new centrifuge tube;
(7) Repeating the steps (4) to (6) by using chloroform/isoamyl alcohol (volume ratio is 24:1);
(8) Adding 0.7 times of pre-cooled isopropanol at-20deg.C, mixing, and standing at room temperature for 10min;
(9) Centrifuging at 12000rpm at room temperature for 15min;
(10) Pouring out the supernatant, washing the precipitate with 500 mu L of 70% ethanol precooled at-20 ℃ for 2-3 times;
(11) After drying the pellet, 50. Mu.L of deionized water or TE was used to dissolve the DNA and placed at-20℃for further use.
(12) 5 μl of the dissolved DNA was extracted, 45 μl of deionized water was added, and the genomic DNA of peanut was prepared by mixing well.
PCR reaction and electrophoresis detection: molecular marker detection is carried out on the parent and all F1 hybrids by utilizing a specific primer pair (shown as SEQ ID No.1 and SEQ ID No. 2) of AhyCs1, and the true hybrid seeds containing male parent and female parent specific strips are obtained according to electrophoresis results.
The PCR amplification reaction system is as follows:
total amplified volume 20. Mu.L:
DNA template 20-30 ng/. Mu.L 1. Mu.L, specific primer pair 0.5 pmol/. Mu.L each 0.5. Mu.L, 10mM dNTP mix 0.5. Mu.L, 10 XTaq Buffer 2.0. Mu.L, 25mM MgCl 2 2.0. Mu.L, taq enzyme 5U/. Mu.L 0.20. Mu.L, and water to 20. Mu.L.
PCR reaction conditions: pre-denaturation at 95℃for 3min;94℃for 30s,56℃for 30s,72℃for 20s,35 cycles; extending at 72℃for 5min.
PCR amplification product detection was performed using 8% non-denaturing polyacrylamide gel (Acr: bis=39:1) electrophoresis.
Wherein, the method for preparing 8% modified polyacrylamide gel is as follows;
to 10. Mu.L of the amplified product was added 3. Mu.L of an indicator-loading buffer (containing 50mM Tris-HCl pH8.0, 50mM EDTA,0.25% bromophenol blue, 0.25% xylene blue, 50% glycerol);
the electrophoresis buffer system was 1 XTBE (90 mM Tris-borate pH 8.3,2mM EDTA), and was electrophoresed at 120V for about 4 hours.
30ml of 8% non-denaturing polyacrylamide gel was formulated as shown in Table 1:
table 130ml 8% non-denaturing polyacrylamide gel formulation
Silver staining detection, the method is as follows:
500mL of 0.1% silver nitrate solution is dyed for 15-20min.
b. Deionized water is quickly rinsed for about 15 sec.
c. Developing solution (1000 mL deionized water +20g NaOH +0.5g Na) 2 CO 3 1.5mL formaldehyde as is) is developed, and shaking is continued until the DNA bands are clearly visible.
d. And (5) rinsing with tap water.
e. Scanning and photographing.
As shown in FIG. 5, the offspring with both male and female parent bands were true hybrids as detected by electrophoresis.
(3) Backcross and offspring screening
With molecular marker assisted recurrent selection breeding protocols, two seasons per year, the entire cycle requires about three years of generation (fig. 6). First, using common high-yield high-quality peanut as female parent (recurrent parent, pink seed coat), true hybrid F 1 Hybridization as male parent, hybridization method is the same as above, harvesting BC 1 F 1 And detecting again by using AhyCs1 molecular markers, and reserving offspring with male parent specific bands, wherein the method for detecting DNA extraction and molecular markers is the same as that described above. Continuously performing backcross and screening for 4 times to obtain BC 4 F 1 And (3) generating, then, carrying out selfing, and selecting pure offspring for seed examination and variety registration.
As the seed coat is developed from the beads, the color of the seed coat is inherited by the parent, and compared with other characters, the color of the seed coat can be displayed in alternate generations. Therefore, the color spot seed husks can survive with great blindness by naked eyes screening and cultivating. By using the method for combining the markers with the backcross recurrent selection, the breeding efficiency can be improved, and the germplasm innovation of the color spot seed coat peanuts can be realized within about 3 years (figure 6). Compared with the traditional method, the efficiency is higher.
The above embodiments are only illustrative of the preferred embodiments of the present invention and are not intended to limit the scope of the present invention, and various modifications and improvements made by those skilled in the art to the technical solutions of the present invention should fall within the protection scope defined by the claims of the present invention without departing from the design spirit of the present invention.
Claims (5)
1. A method for identifying peanut seed coat color by using a specific primer pair of molecular marker AhyCs1, which is characterized by comprising the following steps:
extracting DNA of peanut leaves or seeds to be detected as a template, carrying out PCR (polymerase chain reaction) amplification by using the specific primer pair of the molecular marker AhyCs1, then carrying out gel electrophoresis detection on an amplification product, and judging the seed coat color of the peanut to be detected according to a gel electrophoresis result;
if the gel electrophoresis result only shows a characteristic band with the size of 233bp, judging that seeds of the peanut and the offspring thereof to be detected are obvious color spot streak skin; if the gel electrophoresis result shows that characteristic bands with the sizes of 229bp and 233bp are simultaneously generated, judging that seeds of the peanut and the offspring thereof to be detected are light colored spot striped skins; if the gel electrophoresis result only shows a characteristic band with the size of 229bp, judging that seeds of the peanut to be detected and offspring thereof are not color spot streak skin;
the specific primer pair of the molecular marker AhyCs1 is as follows:
a primer with a nucleotide sequence shown as SEQ ID No. 1;
the nucleotide sequence is shown as a primer shown as SEQ ID No. 2.
2. The method of claim 1, wherein the PCR amplified PCR system is: 1. Mu.L of DNA template, 0.5. Mu.L of forward primer and reverse primer each, 0.5. Mu.L of dNTPMix, 2.0. Mu.L of 10 XTaqBuffer, mgCl 2 2.0. Mu.L of Taq enzyme, 0.20. Mu.L of water was added to 20. Mu.L.
3. The method of claim 1, wherein the PCR procedure for PCR amplification is: pre-denaturation at 95℃for 3min;94℃for 30s,56℃for 30s,72℃for 20s,35 cycles; extending at 72℃for 5min.
4. The method of claim 1, wherein the gel electrophoresis detection comprises 8% non-denaturing polyacrylamide gel electrophoresis.
5. The application of the specific primer pair of the molecular marker AhyCs1 in identifying the peanut with the seed coat color speckle is characterized in that the specific primer of the molecular marker AhyCs1 is used for carrying out PCR amplification on peanut DNA to be detected, then the amplified product is subjected to gel electrophoresis detection, and the seed coat color of the peanut to be detected is judged according to the gel electrophoresis result;
if the gel electrophoresis result only shows a characteristic band with the size of 233bp, judging that seeds of the peanut and the offspring thereof to be detected are obvious color spot streak skin; if the gel electrophoresis result shows that characteristic bands with the sizes of 229bp and 233bp are simultaneously generated, judging that seeds of the peanut and the offspring thereof to be detected are light colored spot striped skins; if the gel electrophoresis result only shows a characteristic band with the size of 229bp, judging that the seeds of the peanut and the offspring thereof to be detected are not the color spot streak skin.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004269487A (en) * | 2003-01-14 | 2004-09-30 | Efuekuto:Kk | Method for production of proanthocyanidin derived from peanut |
| CN113308564A (en) * | 2021-06-30 | 2021-08-27 | 山东省农业科学院 | Molecular marker Ahyrscc closely linked with dark red peanut seed coat and application thereof |
| CN114525364A (en) * | 2022-03-18 | 2022-05-24 | 山东省农业科学院 | Molecular marker AhyWptc tightly linked with peanut white seed coat and application thereof |
| CN115011728A (en) * | 2022-06-28 | 2022-09-06 | 河南省农业科学院 | Molecular marker of peanut endopleura color regulation gene AhLAC function and application |
-
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004269487A (en) * | 2003-01-14 | 2004-09-30 | Efuekuto:Kk | Method for production of proanthocyanidin derived from peanut |
| CN113308564A (en) * | 2021-06-30 | 2021-08-27 | 山东省农业科学院 | Molecular marker Ahyrscc closely linked with dark red peanut seed coat and application thereof |
| CN114525364A (en) * | 2022-03-18 | 2022-05-24 | 山东省农业科学院 | Molecular marker AhyWptc tightly linked with peanut white seed coat and application thereof |
| CN115011728A (en) * | 2022-06-28 | 2022-09-06 | 河南省农业科学院 | Molecular marker of peanut endopleura color regulation gene AhLAC function and application |
Non-Patent Citations (2)
| Title |
|---|
| Transcriptome Analysis of a New Peanut Seed Coat Mutant for the Physiological Regulatory Mechanism Involved in Seed Coat Cracking and Pigmentation;Wan L等;Front. PlantSci.;第7卷;文献号1491 * |
| 花生彩斑调控基因的定位及分子标记开发;马婧;中国知网;全文 * |
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