CN110747288A - Rice large grain gene function marker and application - Google Patents
Rice large grain gene function marker and application Download PDFInfo
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Abstract
The invention discloses a rice large grain gene function marker and application thereof. The functional marker is selected from one or more of GS2 functional marker, GW2 functional marker, GL3.1 functional marker and GW2 promoter marker, wherein the GS2 functional marker is dCaps marker developed aiming at TC/AA mutation of third exon of GS2 gene; the GW2 functional marker is a dCaps marker developed aiming at the nucleotide A deletion mutation of the fifth exon of the GW2 gene; the GL3.1 functional marker is a dCaps marker developed for single base C/A mutation of the tenth exon of the GL3.1 gene; the GW2 promoter marker was the dCaps marker developed for the T/A mutation 858bp upstream of the GW2 translation start site. The invention develops an effective dCaps marker aiming at key variation sites of rice large-grain genes, can effectively identify the rice germplasm of the large-grain rice, and is applied to rice molecular breeding.
Description
Technical Field
The invention relates to rice breeding, in particular to a rice large grain gene function marker and application thereof.
Background
Rice is an important grain crop, and the high yield of the rice determines the grain production safety and social stability. The rice yield is mainly determined by the number of ears, the number of grains per ear and the thousand grain weight. On the premise of not influencing the other two yield factors, the rice yield can be rapidly improved by increasing the thousand seed weight; in addition, in the process of hybrid rice seed production, a large-grain restorer line can be purposefully created, a sterile line is cultivated into a small-grain restorer line, mixed seed and mixed harvest of two lines can be realized, and finally, large-grain and small-grain lines are separated according to grain weight, so that the seed production cost can be greatly reduced. Therefore, the cultivation of large-grain rice varieties has extremely high production and application prospects. However, the current rice breeding still relies on a phenotype observation technology to carry out grain type screening, and although large-grain lines can also be found, the population quantity to be planted is large, and screening can only be carried out after plants are completely mature, and the obtained large-grain lines cannot give consideration to other yield traits. The molecular marker assisted selection technology can identify the development genotype of a plant in the seedling stage of the plant, find the genotype linked with the large-grain character, plant the plant group containing the target genotype when transplanting the rice seedling, almost all the obtained plants are large-grain plant lines when the obtained plants are mature, and the high-yield plant lines with balanced comprehensive characters can be obtained only by considering the characters of the number of ears and the number of grains per ear.
The premise of carrying out large-grain trait molecular screening is to search related germplasm resources and excavate regulatory genes thereof, at present, three large-grain genes are successfully cloned in rice, wherein the gene numbers of the three genes are GS2, GW2 and GL3.1 respectively, and the gene numbers of the three genes are Os02g0701300, Os02g0244100 and Os03g0646900 respectively, wherein GS2 is cloned from a large-grain variety 'Bao-big-grain', and GW2 and GL3.1 are cloned from a large-grain variety 'WY 3'. As shown in fig. 1, the GS2 gene contains five exons, and TC mutation of the third exon of the large grain variety to AA results in the generation of a large grain phenotype; the GW2 gene comprises 8 exons, and the deletion of nucleotide A of the fifth exon of a large-grained variety causes the generation of a large-grained phenotype; the GL3.1 gene comprises 21 exons, and the substitution of a single base C for a in the tenth exon of a large grain variety results in the production of a large grain phenotype. The Sanger sequencing method can be used for directly identifying the key variation, but the sequencing cost is high, one sequencing reaction is 13 yuan in the current market price, and the identification cost of three sites is about 39 yuan. Compared with sequencing, the molecular marker of gene PCR amplification and gel electrophoresis technology has lower cost, but because the three variations belong to base substitution or single base deletion, the PCR amplification products cannot be directly distinguished by gel electrophoresis, and simultaneously, the three variations do not generate enzyme cutting site difference and cannot be identified by utilizing a PCR product enzyme cutting method, no effective molecular marker exists for the three key variations at present, and the molecular breeding improvement of rice large-particle property is greatly limited.
Disclosure of Invention
The purpose of the invention is as follows: in order to solve the problems in the prior art, the invention provides a rice large-grain gene function marker, and provides an effective functional molecular marker for key variation of large-grain genes GS2, GW2 and GL 3.1.
The technical scheme is as follows: the rice large-grain gene function marker is selected from one or more of GS2 function markers, GW2 function markers, GL3.1 function markers and GW2 promoter markers, wherein,
the GS2 functional marker is a dCaps marker developed aiming at TC/AA mutation of a third exon of a GS2 gene;
the GW2 functional marker is a dCaps marker developed aiming at nucleotide A deletion mutation of a fifth exon of GW2 gene;
the GL3.1 functional marker is a dCaps marker developed aiming at the single-base C/A mutation of the tenth exon of the GL3.1 gene;
the GW2 promoter marker is a dCaps marker developed aiming at T/A mutation 858bp upstream of the GW2 translation initiation site.
Wherein the content of the first and second substances,
the GS2 function-labeled primer is as follows:
a forward primer: 5'-AGCGCCACATGCACCGCGGCCGCATACGT-3'
Reverse primer: 5'-TTGCCTGTTCCACCACCAACAGC-3', respectively;
the GW2 function-labeled primers are as follows:
a forward primer: 5'-CACGATACTCCACAGCATAACTGGGAGTCT-3'
Reverse primer: 5'-CTCACACTGCTCAGCCTACA-3', respectively;
the GL3.1 function-labeled primers are as follows:
a forward primer: 5'-TGCACGATTCTATCTGGTTCAGTGGTCGA-3'
Reverse primer: 5'-CTAAACAAACAGGTTTTCTTAC-3', respectively;
the GW2 promoter labeled primer is as follows:
a forward primer: 5'-AAAACCAAAACCTAACACGTGGATACAACA-3'
Reverse primer: 5'-GGCGGTGAAGATAGATGTACT-3' are provided.
The invention also provides application of the rice large grain gene function marker in auxiliary selection of rice grain type or grain weight. Wherein, the GS2 functional marker, the GW2 functional marker and the GL3.1 functional marker can be used for assisting in selecting rice grain types, and the GW2 promoter marker can be used for assisting in selecting grain weights.
The invention also provides a rice breeding method, which is characterized by comprising the following steps: in the breeding process, the functional marker is adopted to assist in selecting the character or grain weight of the rice grain type.
Wherein, the rice grain type is large.
Has the advantages that:
aiming at key mutation sites of rice genes GS2, GW2 and GL3.1, the invention develops an effective dCaps marker, can realize effective amplification and enzyme digestion, can effectively identify large-grain rice germplasm, and is applied to rice molecular breeding.
By utilizing the molecular marker, the grain type and the grain weight traits can be subjected to auxiliary selection, and the breeding process and the purposiveness are accelerated.
The dCaps marker developed by the invention has good primer specificity, and the selected enzymes are common endonuclease with lower price, so that the identification cost can be greatly reduced, and the dCaps marker is suitable for analyzing the genotypes of a large number of groups.
Drawings
FIG. 1 is a schematic diagram of the structure and key variation of three large-grained genes;
FIG. 2 is a schematic diagram of the design of key variant functional molecular markers of the GS2 gene;
FIG. 3 is a schematic diagram of GL3.1 gene key variant functional molecular marker design;
FIG. 4 is a schematic diagram of GW2 gene key variant functional molecular marker design;
FIG. 5 is a schematic diagram of grain types of eight marker-verified varieties;
FIG. 6 is a photograph of GS2 labeled gel electrophoresis;
FIG. 7 shows the gel electrophoresis of GL3.1 marker;
FIG. 8 is a GW2 labeled gel electrophoresis image;
FIG. 9 is a schematic diagram of GW2 promoter SNP marker design;
FIG. 10 shows the GW2 promoter labeled gel electrophoresis.
Detailed Description
The invention will be further elucidated with reference to the following specific examples.
First, function tag development
The gene numbers of the three genes of GS2, GW2 and GL3.1 are Os02g0701300, Os02g0244100 and Os03g0646900 respectively, wherein TC mutation of the third exon of the GS2 gene into AA causes large-grained phenotype generation; the deletion of nucleotide A of the fifth exon of the GW2 gene leads to the generation of a large-grained phenotype; substitution of the tenth exon of the GL3.1 gene for a single base C resulted in the generation of a large-grained phenotype.
Firstly, genome DNA sequences of GS2, GW2 and GL3.1 genes are respectively downloaded from a rice Nipponbare reference genome database to be wild-type sequences, and then, wild-type variants at corresponding positions are respectively replaced by three variant mutants, namely TC of the GS2 gene is replaced by AA, A base of the GW2 gene is deleted, and C of the GL3.1 gene is replaced by A, so that three mutant sequences are obtained. We also confirmed that the mutation types of three genes in two varieties (WY3, Tanbo) were subjected to Sanger sequencing at the same time, and the related mutations were indeed present. We firstly compare the enzyme cutting site difference of the mutant type and the wild type of each gene, and find that the three mutation types can not generate the enzyme cutting site difference, so that the three mutation types can not be developed into Caps labels which are directly cut by enzyme after PCR amplification. Therefore, we have adopted a derived Caps label design approach, where the forward primer is located directly to the left or right of the mutation site, and by introducing 1-2 base mutations into the primer, it can combine with the original mutation to generate a cleavage site difference. Based on the design idea, primer mutation and enzyme cutting site search are carried out by using dCapsFinder2.0 tool, the method is that wild type and mutant type variation and 29-30bp on both sides of the wild type and mutant type variation are copied to corresponding sequence frames, the total 60bp sequence is copied, the number of primer mutation is set to be 2, and a series of primer sequences with mutant bases are obtained after operation. By adopting the method, the forward amplification primers of GS2 and GL3.1 variation are successfully obtained, wherein the forward amplification primer marked by GS2 is AGCGCCACATGCACCGCGGCCGCATACGT, compared with the original sequence, the 25 th base of the primer is replaced by T from A, the 26 th base is replaced by A from C, and a SnaBI enzyme digestion site TACGTA (shown in figure 2) can be formed by combining the primer and the A base of the mutant SNP; the GL 3.1-labeled forward amplification primer is TGCACGATTCTATCTGGTTCAGTGGTCGA, compared with the original sequence, the 25 th base of the primer sequence is replaced by G from C, the 27 th base is replaced by C from A, and a SalI enzyme cutting site GTCGAC can be formed with the C base of the wild-type SNP (FIG. 3). GW2 gene belongs to base deletion mutation type, and cannot use dCapsFinder2.0 tool to search for proper mutation and enzyme cutting site, we directly observe sequence characteristics near the deletion base to search for potential enzyme cutting site, find that TGCAG sequence characteristics exist at the single base deletion position, the recognition sequence of PstI enzyme cutting site is CTGCAG, TTTTGCAG corresponding to wild type variation, and the sequence characteristics after the single base deletion is TTTGCAG, so that the second T can be replaced by C, the wild type sequence becomes TCTTGCAG, the enzyme cutting site still does not exist, the mutant type sequence becomes TCTGCAG, PstI mutation site is generated, and based on the design, the left 30bp sequence is directly selected as forward primer, and the sequence is CACGATACTCCACAGCATAACTGGGAGTCT (figure 4). So far, we successfully obtain three forward primers introduced with specific enzyme cutting sites, and then design reverse paired primers, wherein the design method is to copy the forward primers and 300bp sequences downstream of the forward primers to PremierPrimer 5.0 software, the range of the forward primers is still within the original 30bp interval, the reverse primers are arranged outside the 30bp interval, the highest scoring primer is used as the reverse primer after operation, and finally we respectively obtain a reverse primer sequence CTAAACAAACAGGTTTTCTTAC marked by a reverse primer sequence TTGCCTGTTCCACCACCAACAGC, GL3.1 marked by GS2 and a reverse primer sequence CTCACACTGCTCAGCCTACA marked by GW2, and the forward primers form three pairs of molecular markers (figures 2, 3 and 4). According to the primer position and the enzyme cutting site information, the size of GS2 labeled amplification is 286bp, the size after enzyme cutting is about 258bp, and the mutant site can be subjected to enzyme cutting; the amplification size of the GL3.1 marker is 225bp, the size after enzyme digestion is about 197bp, and a wild type locus can be subjected to enzyme digestion; the amplified size of GW2 marker is 159/158bp, the size after enzyme digestion is about 126bp, and the mutant site can be enzyme digested. The price of the restriction enzymes used by the three markers is relatively low, according to the market price of the three enzymes, the cost of one reaction for identifying the SnaBI enzyme is about 2.7 yuan, the cost of one reaction for identifying the SalI enzyme is about 0.5 yuan, and the cost of one reaction for identifying the PstI enzyme is about 0.16 yuan which is far lower than the price of Sanger sequencing, so that the identification cost can be greatly reduced, and the molecular identification of a large number of germplasms and populations is realized.
Secondly, verifying the amplification enzyme digestion effect of the functional marker
In order to verify the PCR amplification and enzyme digestion effects of the three pairs of markers in the step one, eight varieties, namely YYP1, Nipponbare (NIP), Zhenshan 97(ZS97), Daohuaxiang (DHX), Kasalath (Kasa), WY3, Baodao (BDL) and Katy, are selected and verified, and the grain types of WY3 and Baodao are obviously superior to those of other varieties as shown in FIG. 5. Sampling 8 different rice variety leaves in a seedling stage, putting the leaves into a 2ml centrifugal tube with steel balls, and extracting DNA by adopting a TPS small-amount extraction method, wherein the method comprises the following specific steps: 1. the leaves are vibrated and crushed by a ball mill, 500ul TPS buffer solution is added, and the mixture is placed for 45 minutes at 65 ℃; 2. centrifuging at 12000 speed for 10 min, sucking 300ul of supernatant into a new centrifuge tube, adding isopropanol with the same volume, and standing at room temperature for 45 min; 3. centrifuging at 12000 speed for 10 min to obtain DNA precipitate, pouring out supernatant, and adding 500ul 75% ethanol; 4. 7500 centrifuging for 5 min, removing supernatant, draining, standing at room temperature for 30 min, and adding 100ul double distilled water to obtain DNA solution.
Then, the extracted DNA is taken as a template to respectively carry out PCR amplification on three pairs of primers, and the reaction system is 20ul and comprises 2ul of DNA template, 2ul of PCR buffer (Beijing Dingguo organism), 0.5ul of forward primer, 0.5ul of reverse primer, 0.3ul of Taq polymerase (Beijing Dingguo organism) and 14.7ul of double distilled water. The PCR reaction conditions are as follows: denaturation at 94 ℃ for 3 min, followed by 35 cycles of "denaturation at 94 ℃ for 20 sec-55 ℃ annealing for 30 sec-72 ℃ extension for 20 sec", and final extension at 72 ℃ for 5 min to complete the reaction. The PCR amplification product has almost no difference in band, and can be identified only by further enzyme digestion, the enzyme digestion reaction system is 20ul, including 10ul of PCR product, 0.5ul of endonuclease (the endonuclease is purchased from Thermo Scientific company), 2ul of corresponding buffer, 7.5ul of double distilled water, and 1 hour of enzyme digestion at 37 ℃.
After PCR and enzyme digestion are finished, sucking 10ul of PCR products for agarose gel electrophoresis with the concentration of 3%, carrying out electrophoresis for 20 minutes, then taking a picture, comparing the difference of PCR band types of different varieties, as shown in figures 6, 7 and 8, wherein three markers can amplify clear bands in eight varieties, wherein the GS2 marker has the amplification size of seven variety bands in 8 varieties of 286bp, which cannot be digested, corresponds to wild type variation, only PCR products of precious large-grain varieties can be digested, the band size is about 258bp, corresponds to mutant variation, and is consistent with expectation (figure 6); GL3.1 marks that only the WY3 variety could not be digested in 8 varieties, the band size was 225bp, corresponding to mutant, as expected, while the remaining seven varieties could be digested, with a size of about 197bp, as wild type (FIG. 7); the GW2 marks that the amplified size of seven varieties of bands among 8 varieties is 159bp, which cannot be enzyme-cut, and only the WY3 variety can be enzyme-cut, and the band size is about 126bp, which corresponds to a mutant type (figure 8). Therefore, the three pairs of markers can effectively and accurately distinguish large-particle variation types in the variety, the amplification effect of the three markers is very good, the strips are clear, and the genotype statistics by means of an automatic picture analysis system is facilitated. Thirdly, identifying and analyzing the rice population by the functional marker
The 280 rice varieties distributed all over the world are subjected to mutation type identification by using the three pairs of functional markers in the step (I) and adopting the same amplification enzyme digestion method as the step (II), wherein the mutation type identification comprises six subspecies, namely fragrant rice (AROMATIC), autumn rice (AUS), indica rice (IND), temperate japonica rice (TEJ), tropical japonica rice (TRJ) and mixed group (ADMIX, the genome comprises different subspecies bloods), and the number of corresponding varieties is respectively 10, 43, 58, 55, 65 and 49. The result shows that 280 varieties do not contain three large-grain mutation types, which indicates that the three mutations are not widely applied to rice breeding and have extremely high breeding utilization value, and the three markers designed by the invention can accelerate the application of the three markers in the improvement of large-grain characters of rice.
Fourth, GW2 promoter variation molecular marker is used for identifying and analyzing rice population
Studies have shown that GW2 promoter variation is also associated with the grain weight phenotype, and no single-base deletion type mutation in the GW2 coding region was found in the above 280 varieties, and we speculated that it might contain a promoter region variation that promotes grain weight production, and therefore we decided to further design molecular markers for GW2 promoter region variation. Through a rice polymorphism database website (http:// ricevarmap. ncpgr. cn/v2), the sequence variation of a 2Kb promoter region of GW2 is searched, a large number of SNP variations are found, the SNP with the number of vg0208114649 is screened by comparing and is subjected to marker design, and the database shows that the SNP shows relatively balanced distribution in different subspecies, can be effectively used for genotyping and grain effect evaluation of different subspecies, and is positioned at 858bp upstream of a GW2 translation initiation site. The SNP is a base substitution from T to A, the flanking sequence is CCTAACACGTGGATACAAAA [ T/A ] GCAACCTGGACCCCACGTAA, and for the convenience of primer design, the GW2 promoter sequence is downloaded and the flanking sequence is used for locking the SNP position. The design method of the marker is similar to the three functional markers, namely, a mutation site is introduced into the forward primer to be combined with SNP to generate an enzyme cutting site, and a reverse primer is obtained to be matched with the enzyme cutting site. Firstly, we obtained the forward primer AAAACCAAAACCTAACACGTGGATACAACA of the marker, as shown in fig. 9, compared with the original sequence, the 29 th base of the primer is replaced by a to C, and forms an NspI cleavage site with the T base in SNP, the recognition sequence of the enzyme is RCATGY, wherein R represents a or G, Y represents C or T, the reverse primer is GGCGGTGAAGATAGATGTACT, the PCR amplification size of the marker is 116bp, the size after cleavage is about 86bp, and the reference genome variety nipponica corresponds to the SNP base T, which can be cleaved by enzyme. The PCR amplification and enzyme digestion analysis of DNA of eight varieties, namely YYP1, Nippon (NIP), Zhenshan 97(ZS97), rice flower fragrance (DHX), Kasalath (Kasa), WY3, Bao big particle (BDL) and Katy, are carried out by utilizing the marker, and the result shows that the marker can amplify bands in the eight varieties (figure 10), but the band brightness is slightly weaker than that of the three functional markers, the PCR bands of the YYP1, Kasa and Katy three varieties in the eight varieties are 116bp and cannot be enzyme digested, and the PCR bands of the rest five varieties, namely the Nippon and the like, have the sizes of about 86bp and can be enzyme digested and correspond to T mutation types, so the marker can effectively distinguish T-A variation of 858bp in a GW2 promoter region.
By using the GW2 promoter region identification marker, 280 varieties all over the world are identified, and the result shows that the varieties containing T variation types are 107, the number of the varieties containing A variation types is 169, and 4 varieties are heterozygous, so that the marker can also be used for genotype analysis of hybrid rice and segregating populations. The marker has two genotypes distributed in different subspecies, wherein each genotype of the three subtypes ADMIX, IND and TRJ comprises at least 10 varieties.
TABLE 1 identification of 280 varieties genotype distribution by GW2 promoter marker
Further, we performed genotype-to-grain correlation analysis on three sub-species ADMIX, IND and TRJ. The performance of grain type and grain weight characters is easily influenced by the environment, so that character measurement is carried out in two completely different planting environments of Shanghai and Hainan, after the plants are completely mature, single plant harvesting and drying are carried out on different varieties, then ten thousand-deep SC-G automatic seed test analysis and thousand-grain weight instrument analysis are adopted, grain length, grain width, length-width ratio and thousand-grain weight data are automatically obtained, and then the character difference of two mutant types in subspecies is compared. The specific method comprises the steps of grouping the phenotype data according to two genotypes of each marker, calculating the average value of each group, and carrying out Student' sT-test difference detection, wherein if P is less than 0.05, the marker is considered to be obviously related to the corresponding grain type character. Under the Shanghai planting condition, the two genotypes are found to have no significant influence on grain length and grain width, the T type can increase the length-width ratio in TRJ subspecies, the T type can increase the grain weight in IND subspecies, and although the grain weight difference between ADMIX and TRJ subspecies does not reach a significant level, the grain weight change trend is consistent with that of the IND subspecies, and the T type is proved to have the potential of increasing the grain weight of a plurality of subspecies; under the Hainan planting condition, T-type variation in the IND subspecies can show the grain weight, but the effect in other two subspecies is not obvious, which indicates that the regulation and control of the variation on the grain weight are influenced by genetic background and environmental change, and the marker can effectively eliminate the interference and accurately screen the target genotype.
TABLE 2 identification effect of the GW2 promoter marker on the Shanghai planting conditions
TABLE 3 identification effect of the granule type of GW2 promoter marker under Hainan planting conditions
So far, functional molecular markers of three genes and a grain weight linkage molecular marker are successfully obtained, wherein the three functional markers exist only in large-grain varieties, two large-grain varieties can be used as donors to be hybridized with current production main cultivars and are continuously backcrossed, the molecular marker designed by the invention can accurately screen a plurality of target genes, so that the large-grain properties are rapidly improved, and the T-type grain weight linkage marker can be mainly used for improving indica rice and AUS rice varieties with majority of A mutation types, such as YYP1, Kasalath and Katy varieties used in the invention, the grain weights of the varieties are smaller, and the yield potential can be increased by the marker improvement.
Claims (6)
1. A rice large grain gene function marker is characterized in that the rice large grain gene function marker is selected from one or more of GS2 function marker, GW2 function marker, GL3.1 function marker and GW2 promoter marker, wherein,
the GS2 functional marker is a dCaps marker developed aiming at TC/AA mutation of a third exon of a GS2 gene;
the GW2 functional marker is a dCaps marker developed aiming at nucleotide A deletion mutation of a fifth exon of GW2 gene;
the GL3.1 functional marker is a dCaps marker developed aiming at the single-base C/A mutation of the tenth exon of the GL3.1 gene;
the GW2 promoter marker is a dCaps marker developed aiming at T/A mutation 858bp upstream of the GW2 translation initiation site.
2. The rice large-grain gene function marker according to claim 1,
the GS2 function-labeled primer is as follows:
a forward primer: 5'-agcgccacatgcaccgcggccgcatacgt-3'
Reverse primer: 5'-ttgcctgttccaccaccaacagc-3', respectively;
the gW2 functionally labeled primers are as follows:
a forward primer: 5'-cacgatactccacagcataactgggagtct-3'
Reverse primer: 5'-ctcacactgctcagcctaca-3', respectively;
the primer marked by the gl3.1 function is as follows:
a forward primer: 5'-tgcacgattctatctggttcagtggtcga-3'
Reverse primer: 5'-ctaaacaaacaggttttcttac-3', respectively;
the primer marked by the gw2 promoter is as follows:
a forward primer: 5'-aaaaccaaaacctaacacgtggatacaaca-3'
Reverse primer: 5'-ggcggtgaagatagatgtact-3' are provided.
3. The use of the rice large-grain gene function marker according to claim 1 or 2 in auxiliary selection of rice grain type or grain weight.
4. Use according to claim 3, wherein the rice grain type is large.
5. A rice breeding method is characterized by comprising the following steps: the functional marker of claim 1 or 2 is used to assist in the selection of rice grain type traits or grain weight during breeding.
6. The rice breeding method according to claim 5, wherein the rice grain type is large.
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| CN111876513A (en) * | 2020-06-23 | 2020-11-03 | 湖南杂交水稻研究中心 | Functional marker for detecting nitrogen-efficient and cold-resistant gene OsGRF4 of rice and application thereof |
| CN112609017A (en) * | 2020-12-08 | 2021-04-06 | 浙江大学 | Molecular marker for detecting rice grain shape, corresponding gene and application |
| CN112609018A (en) * | 2020-12-11 | 2021-04-06 | 华智生物技术有限公司 | SNP molecular marker of rice grain type related gene GLW2 and application thereof |
| CN113801956A (en) * | 2021-10-20 | 2021-12-17 | 广西壮族自治区农业科学院 | Fluorescent molecular marker primer group of rice GS2 gene and application thereof |
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| CN112609017A (en) * | 2020-12-08 | 2021-04-06 | 浙江大学 | Molecular marker for detecting rice grain shape, corresponding gene and application |
| CN112609018A (en) * | 2020-12-11 | 2021-04-06 | 华智生物技术有限公司 | SNP molecular marker of rice grain type related gene GLW2 and application thereof |
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