CN111206046B - Transgenic expression vector for regulating grain color of brassica napus as well as construction method and application thereof - Google Patents
Transgenic expression vector for regulating grain color of brassica napus as well as construction method and application thereof Download PDFInfo
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Abstract
The invention belongs to the field of genetic engineering, and particularly relates to a transgenic expression vector for regulating and controlling the grain color of cabbage type rape, a construction method and application thereof, wherein the transgenic expression vector for regulating and controlling the grain color of the cabbage type rape is an RNAi-BnMYB47 transgenic expression vector, the nucleotide sequence of an RNAi fragment of the RNAi-BnMYB47 transgenic expression vector is shown as SEQ ID NO 1, and the transgenic expression vector is prepared by cloning the RNAi fragment of a BnMYB47 gene, then connecting with pFGC5941M plasmid, transforming and extracting plasmid. The RNAi fragment and the expression vector of the BnMYB47 gene provided by the invention can be applied to regulating and controlling the grain color of the cabbage type rape, overcomes the problems that the grain color character of the cabbage type rape is unstable, and stable yellow-seeded rape breeding is difficult to obtain, creates conditions for obtaining a new germplasm material of the cabbage type rape transgenic yellow seed with stable inheritance, and has important guiding significance for rape yellow seed breeding, genetic improvement of rape quality for genetic engineering and breeding practice.
Description
Technical Field
The invention belongs to the field of genetic engineering, and particularly relates to a transgenic expression vector for regulating and controlling the grain color of cabbage type rape as well as a construction method and application thereof.
Background
Rape is one of five oil crops in the world and plays an important role in oil crops in the world. The seeds are used as main harvesting organs of the rape, the seed coat accounts for about 10-20% of the dry weight of the seeds and 20-30% of the dry weight of the cake, and the seed coat is rich in pigments and polyphenol substances and can also seriously affect the oil quality and the feeding value of the cake. Therefore, improving the constitution of brassica napus seeds and increasing the oil content per unit area is one of the main goals of brassica napus breeding. The cabbage type yellow seed rape has the advantages of thin seed coat, high protein and oil content, high cake protein content, low content of toxic substances such as cellulose, pigment and the like, and becomes one of the important targets of rape breeding in recent years.
A large number of researches show that the change of the seed coat color of the cabbage type rape is mainly influenced by important secondary metabolites such as proanthocyanidin, polyphenol, lignin and derivatives thereof, and related genes controlling flavonoid synthetic pathways play an important role in the formation of the seed coat color. Flavonoid substances in seeds of brassica crops mainly comprise kaempferol derivatives, quercetin derivatives, isorhamnetin derivatives and epicatechin glucoside, and the flavonoid is also an important factor influencing the color and luster properties of seed coats. Due to the complexity of these material components, the flavonoid content of different plant seeds is also influenced by genotype and environmental conditions. As one of the largest transcription factors in plants, MYB transcription factor is widely involved in the regulation of various biological processes in plants, and plays a very important role in the growth and development processes of plants. Wherein R2R3-MYB transcription factors play an important role in the formation of the final color of tissues and organs through widely participating in various primary and secondary metabolic processes in the plant phenylpropane-flavonoid metabolic pathway. But the method is lack of natural yellow seed gene resources, has complex grain color inheritance mechanism and unstable characters, causes slow development of inheritance and molecular mechanism research, and is a main difficulty of stable yellow seed rape breeding.
Therefore, aiming at the difficulties, the invention discovers and discloses the biological function of the gene BnMYB47 in the aspect of regulating and controlling the grain color of the cabbage type rape, and determines the flavonoid component in the seed coat of the contrast and the transgenic line in different developmental stages by an LC-MS method, thereby defining the main reason of the color difference of the seed coat; through transcriptome sequencing and gene expression pattern analysis, the correlation that the BnYB 47 participates in regulating and controlling the differences of the brassica napus seed coats is revealed. The BnYB 47 transgenic vector and the construction method thereof provided by the invention have important guiding significance for yellow seed breeding of rape and genetic improvement and breeding practice of rape quality of genetic engineering.
Disclosure of Invention
In view of the above, one of the purposes of the present invention is to provide a RNAi-BnYB 47 transgenic expression vector for regulating the grain color of Brassica napus, which has important guiding significance for yellow seed breeding of Brassica napus, genetic improvement of Brassica napus quality in genetic engineering, and breeding practice.
In order to achieve the purpose, the technical scheme of the invention is as follows:
an RNAi-BnYB 47 transgenic expression vector for regulating grain color of cabbage type rape, wherein the nucleotide sequence of an RNAi fragment of the RNAi-BnYB 47 transgenic expression vector is shown as SEQ ID NO. 1.
Secondly, the second purpose of the invention is to provide a construction method of the RNAi-BnYB 47 transgenic expression vector for regulating and controlling the grain color of the cabbage type rape, and the RNAi-BnYB 47 transgenic expression vector can be accurately obtained through the construction method.
In order to achieve the purpose, the technical scheme of the invention is as follows:
the method for constructing the RNAi-BnYB 47 transgenic expression vector comprises the following steps:
1) Cloning of RNAi fragments of BnYB 47 gene: RNAi primers designed according to BnMYB47 gene and cDNA of oil 821 in cabbage type rape black seed strain are used as templates to carry out PCR reaction to obtain positive clones;
2) And (2) carrying out complete double enzyme digestion on a positive clone of the BnYB 47 gene connected with the interference fragment and a pFGC5941M plasmid by using restriction enzyme, respectively recovering an antisense fragment of the BnYB 47 gene and an open-loop pFGC5941M vector framework after electrophoretic detection, and connecting the recovered fragments overnight by using T4 DNA ligase. The obtained recombinant plasmid is transformed into an escherichia coli competent cell again, and a plasmid of a PCR positive clone bacterium liquid is extracted;
3) Carrying out complete double enzyme digestion on the plasmid obtained in the step 2), respectively recovering a sense fragment and an open-loop antisense fragment after electrophoresis detection and a vector skeleton recombined by pFGC5941M, carrying out connection transformation according to the same method, and carrying out multiple PCR detection on obtained clone germ fluid, wherein a positive clone is an RNAi-BnYB 47 transgenic expression vector.
Further, the forward primer in step 1) contains restriction sites BamHI + AatII, and the reverse primer contains restriction sites XbaI + NcoI.
Further, the nucleotide sequence of the forward primer of the primer in the step 1) is shown as SEQ ID NO. 2.
Further, in the step 1), the nucleotide sequence of the reverse primer is shown as SEQ ID NO. 3.
Further, the temperature for overnight ligation in step 2) was 16 ℃.
The invention also aims to provide application of RNAi fragments of BnMYB47 genes for regulating and controlling the grain color of cabbage type rape.
In order to achieve the purpose, the technical scheme of the invention is as follows:
the RNAi fragment of the BnMY 47 gene is applied to the regulation of the grain color of the brassica napus.
The fourth purpose of the invention provides another application of RNAi fragment of BnYB 47 gene for regulating and controlling grain color of cabbage type rape.
In order to achieve the purpose, the technical scheme of the invention is as follows:
the RNAi fragment of the BnMYB47 gene is applied to rape yellow seed breeding and genetic engineering to improve and breed the rape quality.
The fifth purpose of the invention is to provide the application of the RNAi-BnYB 47 transgenic expression vector for regulating and controlling the grain color of the cabbage type rape.
In order to realize the purpose, the technical scheme of the invention is as follows:
the RNAi-BnYB 47 transgenic expression vector is applied to the regulation of the grain color of the cabbage type rape.
The invention also aims to provide another application of the RNAi-BnYB 47 transgenic expression vector for regulating and controlling the grain color of the cabbage type rape.
In order to achieve the purpose, the technical scheme of the invention is as follows:
the RNAi-BnMYB47 transgenic expression vector is applied to rape yellow seed breeding and genetic improvement and breeding of rape quality in genetic engineering.
The invention has the beneficial effects that: the RNAi-BnYB 47 transgenic expression vector for regulating the grain color of the cabbage type rape can influence the color of the seed coat by regulating lipid metabolism, influencing the accumulation of pigments and polyphenols in the seed coat and regulating flavonoid metabolism. The method for constructing the RNAi-BnMYB47 transgenic expression vector can accurately obtain the expression vector, overcomes the problems that the grain color character of the cabbage type rape is unstable and stable yellow-seeded rape breeding is difficult to obtain by the expression vector, creates conditions for obtaining a new germplasm material of the cabbage type rape transgenic yellow seed with stable inheritance, and has important guiding significance for rape yellow seed breeding, genetic improvement of rape quality in genetic engineering and breeding practice.
Drawings
FIG. 1 control ZY821 and RNAi-BnYB 47 agronomic trait analysis;
FIG. 2 quality trait analysis of control ZY821 and RNAi-BnYB 47;
FIG. 3 phenotypic analysis of control ZY821 and RNAi-BnYB 47 seeds at different stages of development;
FIG. 4 epidermal structural analysis of mature seeds of control ZY821 and RNAi-BnYB 47.
Detailed Description
The examples are given for the purpose of better illustration of the invention, but the invention is not limited to the examples. Therefore, those skilled in the art can make insubstantial modifications and adaptations to the embodiments described above without departing from the scope of the present invention.
EXAMPLE 1 cloning of RNAi fragment of BnMY 47 Gene
RNAi primers are designed according to the result of multiple alignment of the obtained cabbage type rape grain color character BnMY 47 gene sequence (the accession number of ganbank is BnaAnng 36570D), wherein a forward primer contains restriction enzyme sites BamHI + AatII, a nucleotide sequence is shown in SEQ ID NO. 2, a reverse primer contains restriction enzyme sites XbaI + NcoI, and a nucleotide sequence is shown in SEQ ID NO. 3, and cDNA of oil 821 in a black cabbage type rape seed strain is used as a template for amplifying a BnM YB47 gene interference fragment from the cabbage type rape.
The amplification reaction system is shown in table 1, and the PCR reaction amplification procedure is: pre-denaturation at 94 ℃ for 2min → 35 amplification cycles (denaturation at 94 ℃ for 45s → annealing at 58 ℃ for 45s → extension at 72 ℃ for 1 min) → extension at 72 ℃ for 10min. Detecting the reaction amplification product by agarose gel electrophoresis, cutting and recovering the target fragment of the BnMYB47 gene, connecting the cut and recovered target fragment to a pMD19-T vector, transforming escherichia coli JM109 competent cells, delivering a PCR positive clone germ fluid to sample sequencing, and obtaining the nucleotide sequence of the BnYB 47 gene interference fragment shown as SEQ ID NO 1.
TABLE 1BnMYB47 gene interference fragment amplification reaction system
Example 2 construction and characterization of RNAi-BnMY 47 transgenic expression vectors
The plasmid extracted from the positive clone of BnMY 47 gene pMD19-T connected with the interference fragment and the plasmid pFGC5941M are completely double digested by restriction endonuclease NcoI + AatII, the antisense fragment of the BnYB 47 gene and the open-loop pFGC5941M vector framework are respectively recovered after electrophoretic detection, and the recovered fragments are connected at 16 ℃ overnight by T4 DNA ligase. The obtained recombinant plasmid is transformed into competent cells of Escherichia coli JM109 again, and a plasmid of a PCR positive clone bacterium liquid is extracted.
Extracting plasmid of positive clone of BnMY 47 gene pMD19-T connected with interference fragment and plasmid of antisense fragment recombined with pFGC5941 by restriction enzyme BamHI + XbaI, carrying out complete double enzyme digestion on the plasmid, recovering the vector skeleton of sense fragment and open-loop antisense fragment recombined with pFGC5941M after electrophoresis detection, carrying out connection transformation according to the same method, carrying out multiple PCR detection of BAR-sF + BAR-sR, F35S3ND + RNAi-F and RPA3NDR + RNAi-R on obtained clone bacterial liquid, and obtaining the positive clone which is the RNA interference vector of cabbage type rape yellow seed character BnYB 47 gene.
Example 3 regulating action of RNAi-BnMYB47 transgenic plant on cabbage type rape grain color
3.1 investigation of agronomic traits of RNAi-BnYB 47 transgenic plants and control plants and determination of quality traits of RNAi-BnYB 47 transgenic families and control seeds
The RNAi-BnYB 47 transgenic T3 generation family and the black seed strain ZY821 material are sowed at the same time, the seedlings are transplanted, and the cultivation and management method is the same as the field production. After the strains are matured, 10 strains of the strains are selected for testing the seeds according to a rape seed testing method. Respectively recording the character indexes of each plant height (cm), the length (cm) of the main inflorescence, the quantity (number) of siliques of the main inflorescence, the one-time branch height (cm), the one-time branch quantity (number), the length (cm) of the siliques, the thousand seed weight (g) and the like, and carrying out data analysis.
The plant height refers to the length from the cotyledonary node to the highest part of the whole plant and is marked as 'cm'; the length of the main inflorescence is the length from the top end of the main inflorescence to the base of the main inflorescence and is marked as 'cm'; the silique number of the main inflorescence refers to the silique number of more than one full or under-full seed on the main inflorescence, and is marked as 'one'; the length from the first effective branch point to the cotyledonary node is marked as 'cm'; the number of primary branches refers to the number of primary branches with more than one effective silique on the main stem, and is marked as 'one'; the silique length is the average length from the base of the silique to the top of the silique and is reported as "cm"; thousand seeds are the 1000 randomly picked seeds after harvest and after natural air drying (the water content is not higher than 9%), and are marked as 'g'.
Near Infrared Reflectance Spectroscopy (NIRS) scanning is an analytical technique that can rapidly detect seed quality traits without damage to the seeds being tested. Quality traits of RNAi-BnMYB47 transgenic T3 generation and black seed strain ZY821 material seeds, oil content, protein content, high oleic acid, arachidic acid, yellow seed R value, erucic acid, thioglycoside, linolenic acid, linoleic acid, oleic acid and palmitic acid, etc., were measured by using NIR System 6500. 5g of whole rape seed was taken for each sample and placed in a cuvet, and the transgenic RNAi-BnYB 47 was scanned three times, five times (one interval of reflectance spectra 400-2500nm and 2nm) against ZY821, recorded separately, and averaged. The used correction model is developed by rape engineering center in Chongqing city. Finally, the phenotype data obtained from the model are statistically analyzed.
In terms of agronomic traits, except for the length of the silique, the RNAi-BnMYB47 transgenic plants have insignificant difference in agronomic traits such as the number of primary branches, plant height, length of main inflorescence, the number of main inflorescence siliques, thousand kernel weight and primary branch height (figure 1, table 2 and table 3); the near-infrared model is used for analyzing the quality and characters of the transgenic strains RNAi-BnYB 47, and the RNAi-BnYB 47 strains have no significant difference in seed protein content, thioglycoside, arachidonic acid content, linolenic acid content and the like, and have significant difference in yellow seed content, oil content, oleic acid content and linoleic acid content (figure 2, table 4 and table 5). The inhibition of the expression of the BnYB 47 gene is proved to play a certain role in regulating and controlling the oil content, the oleic acid and other quality characters of the brassica napus besides obviously influencing the yellow seed number of the brassica napus.
TABLE 2 analysis of variance of agronomic traits for RNAi-BnMY 47-1
* Correlation at the 0.05 level
TABLE 3 analysis of variance of agronomic traits for RNAi-BnMY 47-2
* Correlation at the 0.05 level
TABLE 4 analysis of variance of seed quality traits for RNAi-BnMY 47-1
* Correlation at 0.01 level
TABLE 5 analysis of variance of seed quality traits for RNAi-BnMY 47-2
* Correlation at 0.01 level
3.2 phenotypic analysis of transgenic lines RNAi-BnMY 47 and control seed development
Seeds of different developmental stages of transgenic pedigrees RNAi-BnYB 47 and black seed ZY821 were observed and photographically recorded by Olympus MVX10 stereoscope.
The seed coat structure of the mature seeds of transgenic inbred RNAi-BnYB 47 and black seed ZY821 was analyzed by SU3500 Scanning Electron microscopy Scanning Electron microscopy.
The RNAi-BnYB 47 seeds and the ZY821 seeds in different development periods are observed by using a stereoscope, and the RNAi-BnYB 47 seeds have similar color in the early stage compared with the control ZY821 seeds, the RNAi-BnYB 47 seeds in the middle and later stages are colored later than the control ZY821 seeds, while the RNAi-BnYB 47 seeds in the later stage have obviously lighter color than the ZY821 seeds, the ZY821 seeds are dark brown black, and the RNAi-BnYB 47 seeds are light yellow (figure 3); meanwhile, the seed coat structures of the mature RNAi-BnMYB47 seeds and the mature ZY821 seeds are analyzed by a scanning electron microscope, and the result shows that a plurality of pores similar to leaf pores are formed in the epidermis of the mature seeds, and compared with the control ZY821 seeds, the diameters of the pores on the surfaces of the mature seeds of RNAi-BnYB 47 are smaller, and the cells are compact (figure 4). The observation result of combining a stereoscope and a scanning electron microscope can show that the BnYB 47 gene can participate in the forming process of regulating and controlling the color of the seed coat of the cabbage type rape, the regulation and control degrees in different periods are inconsistent, the BnYB 47 gene can directly participate in the forming process of regulating and controlling the color of the seed coat of the cabbage type rape, and the BnYB 47 gene can be used as a regulation and control factor to regulate and control the color of the seed coat of the cabbage type rape by regulating and controlling one or more genes in the pigment synthesis process. In addition, the hole diameter of the mature RNAi-BnYB 47 seed surface is reduced, the cells are compact, and the inhibition of the expression of BnYB 47 genes can influence the structure of seed coats.
3.3 transgenic lines RNAi-BnMYB47 and control seed coat flavonoid component LC-MS analysis
3.3.1 extraction of Total Flavonoids from transgenic pedigree RNAi-BnMY 47 and control seed coat
(1) Fully freezing and drying fresh seed coat materials in different development stages, loading into an EP tube containing stainless steel balls, and fully oscillating and grinding on a high-speed oscillator;
(2) Weighing 5mg of the fully ground powder into a 5ml glass tube, adding a 1ml mixed solvent of methanol: acetone: water: trifluoroacetic acid (TFA) (40;
(3) Performing ultrasonic disruption treatment at 4 deg.C for 15min, centrifuging at 19000 Xg for 5min, sucking supernatant to clean 2ml EP tube, and storing the obtained sample supernatant at-80 deg.C;
(4) Adding 1ml of a mixed solvent of methanol, acetone, water, trifluoroacetic acid (TFA) (40;
(5) Centrifuging at 19000 Xg for 5min, sucking supernatant, mixing the two supernatants, storing at-80 deg.C in refrigerator, waiting for LC-MS analysis, and storing the precipitate at-80 deg.C for subsequent determination of proanthocyanidin content.
3.3.2 transgenic pedigree RNAi-BnMYB47 and control seed coat flavonoid component LC-MS analysis
LC-MS analysis in Dionex Ulti Mate TM 3000UHPLC systems (Thermo Fisher Scientific, USA), the method can be referred to (Auger et al, 2010), and the specific method is as follows:
(1) All samples were filtered through nylon membranes (0.22 μm, memberan, GER) before analysis, and then 200 μ l of each sample was pipetted into a clean sample tube;
(2) Each sample was sampled in an amount of 5. Mu.l and was separated on an ACQUITY UPLC BEH C18 column (2.1X 150mm,1.7 μm; waters, USA) before entering the mass spectrometer, eluent A was 0.1% acetonitrile formate. The elution gradient is 5-9% A,5min;9% by weight of A-16% by weight of A,10min;16% by weight of A-50% by weight of A,25min;50% of A to 95% of A,15min;95% A,5min, followed by column washing and rebalancing;
(3) Mass spectrometry in Thermo Scientific TM Q-Exactive TM Performed on a mass spectrometer equipped with a HESI source and operated in negative ion mode. The source working parameters are spraying voltage, 3 kV; sheath gas flow rate, 35 units; flow rate of auxiliary gas, 10 units; capillary temperature, 350 ℃; the temperature of an auxiliary gas heater is 300 ℃; nitrogen is used as sheath gas and auxiliary gas;
(4) The major flavonoids were first identified by full MS mode analysis and then MS2 analysis was performed on the selected m/z ions. Data acquisition and subsequent processing analysis used Thermo Xcalibur 3.0.63 (Thermo Fisher Scientific). The flavan-3-ol and soluble PAs extract (procyanidin monomer consisting of flavan-3 alcohol) content is expressed as epicatechin equivalents, relative to the calibration curve for epicatechin, i.e., equivalent μ g of epicotechin per mg of dry weight (μ g EC equiv/mg DW). The content of other phenolic compounds is finally expressed as relative content to the corresponding calibration curve of available standards including p-coumaric acid (p-coumaric acid), caffeic acid (caffeic acid), ferulic acid (ferulic acid), epicatechin (epicatechin), quercetin (quercetin), kaempferol (kaempferol) and isorhamnetin (purchased from PureChem-Standard co., ltd, chengdu, china), as well as sinapic acid (sinapic acid) and coniferyl aldehyde (purchased from Sigma-Aldrich).
LC-MS is utilized to determine the flavonoid components in the seed coats of the control ZY821 and the transgenic line RNAi-BnMYB47 in different development periods, and 27 main flavonoid components are detected in total, wherein the main flavonoid components mainly comprise secondary metabolites such as epicatechin, kaempferol, isorhamnetin, anthocyanin and derivatives thereof (Table 6). Indicating that BnYB 47 may be involved in the synthesis of these metabolites.
TABLE 6 LC-MS identification of major flavonoid component in seed coat during Brassica napus seed development
Finally, the above embodiments are only for illustrating the technical solutions of the present invention and not for limiting, although the present invention has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions may be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention, and all of them should be covered in the claims of the present invention.
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Claims (7)
1. The method for constructing the RNAi-BnYB 47 transgenic expression vector for regulating and controlling the grain color of the cabbage type rape is characterized by comprising the following steps of:
1) Cloning of RNAi fragment of BnYB 47 gene: RNAi primers designed according to BnMYB47 genes and cDNA of oil 821 in the cabbage type rape black seed strain are used as templates to carry out PCR reaction to obtain positive clones; the nucleotide sequence of RNAi fragment of BnMY 47 gene is shown in SEQ ID NO 1;
2) Carrying out complete double enzyme digestion on a positive clone of a BnMY 47 gene connected with an interference fragment and a pFGC5941M plasmid by using restriction enzyme, respectively recovering an antisense fragment of the BnYB 47 gene and an open-loop pFGC5941M vector skeleton after electrophoretic detection, connecting the recovered fragments overnight by using T4 DNA ligase, re-transforming escherichia coli competent cells by using the obtained recombinant plasmid, and extracting a PCR positive clone bacterium liquid plasmid;
3) Carrying out complete double enzyme digestion on the plasmid obtained in the step 2), respectively recovering a sense fragment and an open-loop antisense fragment after electrophoresis detection and a vector skeleton recombined by pFGC5941M, carrying out connection transformation according to the same method, and carrying out multiple PCR detection on obtained clone germ fluid, wherein a positive clone is an RNAi-BnYB 47 transgenic expression vector.
2. The method according to claim 1, wherein the PCR reaction in step 1) is carried out using a forward primer containing the restriction sites BamHI + AatII and a reverse primer containing the restriction sites XbaI + NcoI.
3. The method according to claim 2, wherein the nucleotide sequence of the forward primer for performing the PCR reaction in step 1) is shown as SEQ ID NO. 2.
4. The method according to claim 2, wherein the nucleotide sequence of the reverse primer for the PCR reaction in step 1) is shown in SEQ ID NO. 3.
5. The method of claim 1, wherein the overnight ligation temperature in step 2) is 16 ℃.
An application of RNAi fragment of BnMYB47 gene in regulation of cabbage type rape grain color, wherein the nucleotide sequence of the RNAi fragment is shown as SEQ ID NO. 1.
And 7, application of the RNAi-BnYB 47 transgenic expression vector in regulation of the grain color of the cabbage type rape, wherein the nucleotide sequence of an RNAi fragment of the RNAi-BnYB 47 transgenic expression vector is shown as SEQ ID NO. 1.
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| CN101942455A (en) * | 2010-09-15 | 2011-01-12 | 西南大学 | TT16 gene families of brassica napus and parental species celery cabbage and cabbage of same as well as application thereof |
| CN101942458A (en) * | 2010-09-15 | 2011-01-12 | 西南大学 | Gene families of cabbage type rape, parental species Chinese cabbage and cabbage AHA10 thereof and applications thereof |
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| CN111235159A (en) * | 2018-11-27 | 2020-06-05 | 西南大学 | Application of MYB61 gene in cabbage type rape grain color breeding |
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| CN101942455A (en) * | 2010-09-15 | 2011-01-12 | 西南大学 | TT16 gene families of brassica napus and parental species celery cabbage and cabbage of same as well as application thereof |
| CN101942458A (en) * | 2010-09-15 | 2011-01-12 | 西南大学 | Gene families of cabbage type rape, parental species Chinese cabbage and cabbage AHA10 thereof and applications thereof |
| CN111235159A (en) * | 2018-11-27 | 2020-06-05 | 西南大学 | Application of MYB61 gene in cabbage type rape grain color breeding |
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