CN109810182B - BnLAX1.c gene, protein and application thereof in controlling cabbage type rape plant type - Google Patents
BnLAX1.c gene, protein and application thereof in controlling cabbage type rape plant type Download PDFInfo
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- CN109810182B CN109810182B CN201910092178.6A CN201910092178A CN109810182B CN 109810182 B CN109810182 B CN 109810182B CN 201910092178 A CN201910092178 A CN 201910092178A CN 109810182 B CN109810182 B CN 109810182B
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Abstract
The invention relates to BnLAX1.c gene, protein and application thereof in controlling cabbage type rape plant type, and the invention separates and applies a gene containing BnLAX1.c geneBnLAX1.cA DNA fragment of a gene conferring an increased number of main stems on brassica napus. Wherein said compound containsBnLAX1.cThe nucleotide sequence of the gene coding region is shown as SEQ ID NO. 1 of the sequence table, the sequence length is 534 bp, the amino acid sequence of the coded protein is shown as SEQ ID NO. 2, and the number of the amino acids is 177. The invention relates to the separation of Brassica napusBnLAX1.cThe gene and the identification of the biological function of the gene in the aspect of improving the cabbage type rape plant type have very important significance for cultivating a new variety of high-yield cabbage type rape.
Description
Technical Field
The invention relates to the field of cabbage type rape genetic engineering. In particular to a cabbage type oil which can increase the number of main stems and is obtained by separating, cloning and functional verificationVegetable dishBnLAX1.cApplication of the gene in cabbage type rape plant type genetic improvement. The invention adopts a RT-PCR method to separate the gene for controlling the main stem number of the cabbage type rape BnLAX1.cOver-expression ofBnLAX1.cThe gene can add the number of main stems of the cabbage type rape, and the function and the application way of the gene are proved.
Background
Cabbage type rape (C)Brassica napusL.) is the main rape cultivation species in China, and has the characteristics of high yield, strong stress resistance, wide adaptability and the like. Among the constitutive factors of the yield of the brassica napus, the contribution of one-time effective branches and the quantity of siliques thereof is the largest. The main stem of the cabbage type rape is crucial to the whole growth and development process of the rape, the main stem not only plays a role in supporting and conducting, but also can directly influence the yield of the cabbage type rape by influencing factors such as the number of effective branches at one time, the number of siliques of main inflorescence and the like; cabbage type rape usually has only one main stem, and the multi-main-stem trait can improve the yield-increasing potential of cabbage type rape, so that the trait becomes one of the target traits for Genetic improvement of cabbage type rape (Zhang Y, Li Q, Cui Y, et alBrassica napusL, the therapeutic and Applied Genetics, 2018, 131(11): 2311-2319). Pradhan et al found that the number of primary branches, the number of secondary branches, the total number of siliques per plant, silique density and the like contributed most to Heterosis of yield traits in brassica juncea (Pradhan A K, Sodhi Y S, Mukhopadhyay A, et al Brassica juncea, L. Czern &Coss) Analysis of component reagents to biology for yield Euphytoica, 1993, 69(3): 219-229). The utilization of heterosis to obtain hybrid progeny with good characters is one of effective ways to improve the yield of rape, but the time is too long. The cabbage type rape with obviously increased main stem number obtained by utilizing a transgenic technology is not reported at present.
Disclosure of Invention
In order to overcome the defects, the invention provides a BnLAX1.c gene, a protein and application thereof in controlling the plant type of brassica napus. Selecting one according to the result of RNA-seqA candidate gene specifically expressed in meristems, which the applicant namedBnLAX1.c. The invention separates and uses aBnLAX1.cA DNA fragment of a gene, which fragment confers an ability to increase the number of main stems of Brassica napus. Wherein said compound containsBnLAX1.cThe nucleotide sequence of the gene coding region is shown as SEQ ID NO. 1 of the sequence table, the sequence length is 534 bp, the amino acid sequence of the coded protein is shown as SEQ ID NO. 2, and the number of the amino acids is 177.
In order to achieve the purpose of the invention, the technical scheme adopted by the invention is as follows: a cabbage type rape plant type regulatory protein is characterized in that the protein is (a) or (b) as follows:
(a) A protein consisting of an amino acid sequence shown as SEQ ID NO. 2;
(b) a protein which is obtained by substituting and/or deleting and/or adding one or more amino acid residues to the amino acid sequence of the SEQ ID NO. 2, is related to the plant type regulation of the Brassica napus and is derived from the SEQ ID NO. 2.
It is another object of the present invention to provide a gene encoding the aforementioned protein.
The gene is a DNA molecule of any one of the following (a 1) - (a 3);
(a1) 1, DNA molecule shown in SEQ ID NO;
(a2) a DNA molecule which hybridizes under stringent conditions with the DNA sequence defined in (a 1) and encodes a protein involved in the plant type regulation of Brassica napus;
(a3) a DNA molecule which has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% homology with the DNA sequence defined in (a 1) and encodes a protein involved in the regulation of Brassica napus plant type.
Another object of the present invention is to provide an expression cassette, a recombinant vector, a recombinant microorganism or a transgenic cell line containing the aforementioned gene.
It is a further object of the present invention to provide the use of (b 1) or (b 2) or (b 3) or (b 4):
(b1) The protein, or the gene, or an expression cassette, a recombinant vector, a recombinant microorganism or a transgenic cell line containing the gene is applied to the regulation of the cabbage type rape plant type;
(b2) the protein, or the gene, or an expression cassette, a recombinant vector, a recombinant microorganism or a transgenic cell line containing the gene is applied to culturing a new variety of the brassica napus;
(b3) the protein, or the gene, or an expression cassette, a recombinant vector, a recombinant microorganism or a transgenic cell line containing the gene is applied to the regulation of the number of main stems of the brassica napus;
(b4) the protein, or the gene, or an expression cassette, a recombinant vector, a recombinant microorganism or a transgenic cell line containing the gene is applied to the regulation of the number of main stems of plants.
The invention also provides a method for cultivating the transgenic plant for increasing the number of main stems of the plant, which aims to improve the content or activity of the protein in the target plant to obtain the transgenic plant; the transgenic plant has a higher number of plant main stems than the target plant.
A method for cultivating transgenic plants with increased main stem number comprises cultivating transgenic plants by the method; the transgenic plant expresses cabbage type rape plant type regulatory protein or contains the gene.
Carrying the inventionBnLAX1.cExpression vectors for genes can be introduced into Plant cells by conventional biotechnological methods using Ti plasmids, Plant viral vectors, direct DNA transformation, microinjection, electroporation, etc. (Weissbach, 1998, Method for Plant Molecular Biology VIII, academic Press, New York, pp.411-463; Geiserson and Corey, 1998, Plant Molecular Biology (2nd Edition)).
May use the inventionBnLAX1.cThe expression vector of the gene is used for transforming a host (various plants including cabbage type rape) and cultivating a plant variety with improved plant type. The plant host can also be rice, tobacco, soybean, tomato, wheat, etc.
The gene of the invention has higher expression level in the apical meristem of the plant, so the gene of the invention can be combined with any promoter specifically expressed by the meristem and then connected into a proper expression vector, and a plant host is transformed, so that the number of the meristems of the plant is increased, and the plant type of the plant is changed.
Recovering DNA using a DNA recovery kit comprisingBnLAX1.cThe DNA fragment of the gene coding region is connected into a pMDC83 skeleton vector by utilizing an enzyme digestion connection method to construct an over-expression vector of the gene, which is named as pMDC83-BnLAX1.c。
pMDC83- BnLAX1.cThe vector is introduced into Agrobacterium tumefaciens, and the Agrobacterium tumefaciens strain is GV 3101. pMDC83-BnLAX1.cThe cabbage type rape receptor material J9712 is successfully obtainedBnLAX1.cCompared with the wild type transgenic plant, the transgenic plant with the gene expression quantity remarkably improved has the observation that the overexpression is compared with the wild type plantBnLAX1.cThe number of main stems of the transgenic cabbage type rape is increased, which shows thatBnLAX1.cCan regulate and control plant types.
In conclusion, the invention takes the cabbage type rape as the research material, and discovers a candidate gene specifically expressed in the meristem by analyzing the result of RNA-seq of each tissue organ of the cabbage type rape variety 'A9712', the gene may play an important role in the process of formation and differentiation of the apical meristem, and the gene is named asBnLAX1.c。
Cabbage type rape is an important raw material of edible oil, and the improvement of the yield and the improvement of the quality of the edible oil are always the targets of people's efforts. In the present invention, overexpressionBnLAX1.cThe gene increases the main stem number of the cabbage type rape, which shows thatBnLAX1.cThe gene participates in the plant type regulation of the cabbage type rape. Thus, isolation from Brassica napusBnLAX1.cThe gene and the identification of the biological function of the gene in the improvement of the cabbage type rape plant type have very important significance for cultivating high-yield new cabbage type rape varieties.
Drawings
SEQ ID NO 2 of the sequence Listing isBnLAX1.cAn amino acid sequence of a gene-encoded protein;
FIG. 1 shows a schematic view of aBnLAX1.cThe expression condition of the gene in each tissue organ of the brassica napus;
FIG. 2BnLAX1.cOverexpression vector construction scheme;
FIG. 3BnLAX1.cIn over-expressed plantsBnLAX1.cThe expression of the gene, CK1 and CK2 are negative seedlings of the transgene; BnLAX1-2, BnLAX1-4, BnLAX1-10, BnLAX1-11, BnLAX1-12, BnLAX1-18 areBnLAX1.cTransgenic positive brassica napus;
FIG. 4BnLAX1.cPlant type performance of overexpressing transgenic brassica napus, in the figure: WT is wild type control plant;BnLAX1.c-OE is overexpressedBnLAX1.cThe transgenic brassica napus of (1).
Detailed Description
The following examples define the invention and describe that the invention comprisesBnLAX1.cDNA fragments of the complete coding segment of the gene, and validationBnLAX1.cMethods of gene function. From the following description and these examples, one skilled in the art can ascertain the essential characteristics of this invention, and without departing from the spirit and scope thereof, can make various changes and modifications of the invention to adapt it to various usages and conditions.
Example 1: RNA-seq analysis of endogenesis in Brassica napusBnLAX1.cExpression of genes in various tissues and organs
Tissue and organ samples of the cabbage type rape line 'A9712' at each period are taken, quickly placed in liquid nitrogen for quick freezing, and transferred to a refrigerator at 70 ℃ for storage until RNA is extracted. The total RNA extraction adopts the RNAioso Plus kit of the company TaKaRa, the RNA-Seq analysis is carried out by the Meiji biological medicine science and technology Limited company of Shanghai, and the FPKM value of the gene is analyzed according to the high-throughput sequencing result, so as to extract the total RNABnubi(BnaA 10g 06670D) as reference GeneBnLAX1.cThe relative expression levels in the respective tissues are shown in FIG. 1.
Example 2: brassica napusBnLAX1.cMolecular cloning of genes
Taking three-leaf one-heart-stage seedlings of the cabbage type rape variety Darmor-bzh, quickly freezing the seedlings by liquid nitrogen, and storing the seedlings in a refrigerator at the temperature of 70 ℃ below zero for extracting total RNA. Total RNA was extracted using the RNAioso Plus kit from TaKaRa, Inc. Synthesis of cDNA for Brassica napus first Strand Synthesis was performed according to the HiScript 1st Strand cDNA Synthesis Kit of Nanjing Novowed Biotech, Inc.
Taking the first strand of cDNA synthesized by the kit as an amplification template, and taking the designed F: 5'-ACACAGACACCACGTAACAATAC-3' (SEQ ID NO: 3) and R: 5'-GGGGACACTAACAAACTAAGAT-3' (SEQ ID NO: 4) as primers, and performing cDNA amplification by RT-PCR under the following conditions: 3 min at 94 ℃, 15 s at 59 ℃ and 30 s at 72 ℃ for 35 cycles; 10 min at 72 ℃. And (3) carrying out electrophoretic analysis after the PCR is finished, and recovering the target amplified fragment by adopting a DNA recovery kit of Kangji biological technology limited company. Connecting the amplified fragment with pEASY-Blunt T vector of Beijing Quanji Biotechnology Limited, transforming competent cells of Escherichia coli, selecting white colony to perform colony PCR to identify positive clone, sending the positive clone to Yangzhou Ongke Biotechnology Limited for sequencing, and naming the plasmid without errors after sequencing verification as the plasmid BnLAX1.c-T。
Example 3:BnLAX1.cconstruction of Gene overexpression vector
In order to better analyzeBnLAX1.cThe function of the gene, which the applicant overexpresses in Brassica napus, was studied by observing the phenotype of the transgenic plants.
The construction method of the over-expression vector comprises the following steps: verified by the above sequencingBnLAX1.cGene cloning vector plasmidBnLAX1.c-T as template, sequence specific primer plus linker using primer LAX1F (5'-gACTAGTcATGGACCAGTCCACTCTTCA-3') (SEQ ID NO: 5)SpeI site) and LAX 1R (5'-atGGCGCGCCatAGACAAACCACGTCTATGCA-3') (SEQ ID NO: 6), sequence specific primers plus linkerAscI site)Performing cDNA amplification by using RT-PCR under the following conditions: 3 min at 94 ℃, 15 s at 58 ℃ and 30 s at 72 ℃ for 35 cycles; 10 min at 72 ℃. And (3) carrying out electrophoretic analysis after the PCR is finished, and recovering the target amplified fragment by adopting a DNA recovery kit of Kangji biological technology limited company. Connecting the amplified fragment to pEASY-Blunt T vector of Beijing holotype gold biotechnology, Inc., transforming escherichia coli competent cells, selecting white colony to perform colony PCR to identify positive clone, and sending the positive clone to Yangzhou Ongke Biotechnology, Inc. for sequencing.
IncludedBnLAX1.cCloning vector plasmid of gene fragmentSpeI＋AscAfter double enzyme digestion, the DNA recovery kit is used to recover the target DNA fragment, and the fragment is connected with the pMDC83 skeleton vector of the corresponding enzyme digestion to constructBnLAX1.cThe over-expression vector of the gene was named pMDC83-BnLAX1.c(FIG. 2).
Example 4: pMDC83-BnLAX1.cGenetic transformation of brassica napus with overexpression vector
pMDC83-BnLAX1.cThe plasmid was introduced into competent cells of Agrobacterium tumefaciens GV3101 strain. Selecting single colony, inoculating into 25 mL YEB culture medium (containing 50 mg/L rifampicin), culturing overnight, inoculating 5 mL bacterial liquid into 100 mL YEB culture medium (containing 50 mg/L rifampicin), and culturing to OD600= 0.7-0.8, the bacterial solution is placed on ice for 10 min, centrifuged at 5000 rpm and 4 ℃ for 10 min to collect the thalli, and then 100 mL of sterile double distilled water is added to wash twice. The cells were suspended in 4 mL of 10% glycerol and transferred to a 50 mL centrifuge tube. The strain is collected by centrifugation at 5500 rpm and 4 ℃ for 10 min, 500 mu L of 10% glycerol is added to resuspend the strain, and the strain is transferred to a 1.5 mL centrifuge tube.
Taking 50 mu L competent cells, adding 5 mu L pMDC83-BnLAX1.cThe recombinant plasmid is mixed evenly by a gun head and transferred into an electric transformation cup of 0.1 cm. Electrical conversion parameters: 200 omega, 1.7 KV, 2.5F, and 500 muL of LB culture solution is added immediately after electric shock. After culturing at 37 ℃ and 220 rpm for 1 h, 100 muL of bacterial liquid is taken and coated with LA culture medium containing Kanamycin Kanamycin resistance to screen transformants, and culturing is carried out for 16 h at 28 ℃.
The genetic transformation method for transforming the cabbage type rape is a method after improvement of a transformation method in key laboratories of the national crop genetic improvement of Huazhong agricultural university, takes the hypocotyl of a sterile seedling of the cabbage type rape as an explant, and utilizes an agrobacterium-mediated method to realize the genetic transformation of an exogenous fragment in the cabbage type rape.
The formula of the culture medium is as follows:
inoculation Medium (M)0）：MURASHIGE &SKOOG MEDIUM (Duchefa Biochemie company) +30.0 g/L Sucrose Surose +8 g/L Agar (pH 5.8-pH 6.0).
Co-cultivation Medium (M)1）：M0+ 18.0 g/L Mannitol Mannitol + 1.0 mg/L2, 4-dichlorophenoxyacetic acid 2,4-D +0.3 mg/L Kinetin Kinetin + 100. mu.M acetosyringone AS (pH 5.8).
Callus differentiation medium (M)2）：M1+300.0 mg/L Timentin +25 mg/L Hygromycin B.
Seedling culture medium (M)3）：MURASHIGE &SKOOG MEDIUM (Duchefa Biochemie company) +10.0 g/L Glucose +0.25 g/L Xylose xylosyl +0.6 g/L morpholine ethanesulfonic acid MES +2.0 mg/L Zeatin +0.1 mg/L indoleacetic acid IAA +300.0 mg/L Timentin +25 mg/L Hygromycin B.
Strong seedling rooting culture medium (M)4）：M0+300.0 mg/L Timentin.
Murashige & Skoog media is simply referred to as MS MEDIUM.
The specific operation steps are as follows:
(1) and (3) sterilization:
a. firstly, soaking cabbage type rape seeds in 75% alcohol for 1 min, wherein attention cannot be paid for too long time;
b. then sterilizing with 2% sodium hypochlorite for 20 min;
c. finally, the seeds are washed for 4-5 times by sterile water, and are cleaned as far as possible.
(2) Sowing:
a. seeding the sterilized seeds to M with sterile forceps 030 grains per dish on the culture medium;
b. placing the inoculated culture tank into an incubator, and culturing at 24 ℃ in the dark for 6-7 d.
(3) Shaking the bacteria:
after 5-6 days of sowing, inoculating the agrobacterium into a sterile triangular flask or a centrifuge tube containing LB liquid culture medium, and placing the sterile triangular flask or the centrifuge tube into a shaker at 28 ℃ and 180 and 220 rpm for culturing.
(4) Explants were prepared and infected:
a. cutting seedling growing for 6-7 days after seeding with sterile forceps and scalpel, and cutting hypocotyl into explant segment with length of 0.8-1.0 cm. When cutting seedling, the hypocotyl is arranged in M1The liquid culture medium is cut, so that the cutting effect is better. The cutting is fast and accurate without dragging;
b. measuring OD of Agrobacterium600Value (OD in LB medium)600= about 0.3 is preferable), the previously cultured bacterial solution is centrifuged at 6000 rpm for 10 min, the supernatant is discarded, the bacterial solution is resuspended in an MS liquid medium containing 100. mu.M acetosyringone AS, which is the same volume AS the bacterial solution, and the process is repeated once more. Finally, taking 2 mL of bacterial liquid, and diluting the bacterial liquid with 20 mL of MS liquid culture medium containing 100 mu M acetosyringone AS;
c. And (3) putting the cut explants into the bacterial liquid resuspension with the adjusted concentration, and carrying out dip dyeing for 10 min, wherein the infection time is not too long, otherwise, the explants die. 150-200 explants are suitably impregnated in each 20 mL of the bacterial solution;
(5) transfer of infected explants to M1Culturing on culture medium at 24 deg.C in dark for 36-48 hr with 20-25 explants per dish;
(6) explant from M1Go to M2On the culture medium, and transferred to a light incubator (24 ℃ for 16 h/8 h) for 3 weeks;
(7) transfer of explants to M3Subculturing on the culture medium every 2-3 weeks until green buds appear;
(8) finally transferring the explants to M4Rooting in culture medium for 2-4 weeks.
Example 5: identification of transgenic cabbage type rape positive plants
The rapid method is adopted to extract the genome DNA of the cabbage type rape, and the steps are as follows:
(1) taking two young leaves (about 0.2 g), shearing into pieces, putting into a 2 mL centrifuge tube, adding 250 mu L DNA buffer and two steel balls (diameter is 6.7 mm), and breaking a leaf sample by a sample making machine at 50 Hz for 180 s;
(2) incubating the fragmented sample at 95 ℃ for 10 min;
(3) taking out the sample, cooling to room temperature, and centrifuging at 12000 rpm for 5 min;
(4) And (4) sucking 50 muL of supernatant, transferring the supernatant into a new 1.5 mL centrifuge tube, and diluting by 5 times for later use.
DNA buffer formulation:
Tris-HCl (pH=7.5) 500 mM
NaCl 300 mM
sucrose 300 mM
1 μ L of DNA was used as a template, and PCR amplification was performed with primers 35S (5'-TCCCACTATCCTTCGCAAG-3') (SEQ ID NO: 7) and R (5'-atGGCGCGCCatAGACAAACCACGTCTATGCA-3') (SEQ ID NO: 8), R (5'-gACTAGTcATGGACCAGTCCACTCTTCA-3') (SEQ ID NO: 9) and GFP (5'-TCCCACTATCCTTCGCAAG-3') (SEQ ID NO: 10) under the following conditions: 5 min at 94 ℃; 30 cycles of 94 ℃ for 30 s, 58 ℃ for 30 s and 72 ℃ for 30 s; 10 min at 72 ℃. By using transgenic cabbage type rape DNA as a template, a specific target segment can be amplified, and the target vector is proved to be 35S:BnLAX1.chas been integrated into the genome of Brassica napus.
Example 6: over-expressionBnLAX1.cThe gene increases the number of main stems of transgenic cabbage type rape
The invention adopts a fluorescence detection real-time quantitative method to detect partial transgenic cabbage type rape plantsBnLAX1.cGene expression was detected, RNA was extracted and reverse transcribed as described in example 3. The fluorescent quantitative PCR was performed on 7500 quantitative PCR instrument of ABI company, using qG 1F (5'-CATCTTCTTCTTTACACAGCCG-3') (SEQ ID NO: 11) and qG 1R (5'-ACCAGATCTGAGCTTTCAAGAA-3') (SEQ ID NO: 12) as primers, and the reaction conditions were: 9 Collecting fluorescence signals at 5 ℃ for 1 min, 95 ℃ for 15 s, 60 ℃ for 20 s, and 72 ℃ for 31 s for 40 cycles; fluorescence signals were collected every 1 ℃ for 1 s from 60 ℃ to 95 ℃. After the reaction was completed, analysis and mapping were performed by ABI7500 Software (7500 Software v2.0.1) attached thereto. The result shows that the success is achievedBnLAX1.cExpression level of Gene relative to wild typeBnLAX1.cTransgenic plants with significantly increased gene expression levels (fig. 3).
For over-expressionBnLAX1.cThe transgenic plants of (a) were phenotypically analyzed and found to be over-expressed compared to wild-type control plantsBnLAX1.cThe number of main stems of the transgenic plants was increased (FIG. 4).
The invention separates and obtains the cabbage type rape gene related to the plant type, can pertinently obtain the candidate gene for regulating and controlling the plant type of the plant, and has certain theoretical guidance function for researching the plant branching mechanism and separating the gene related to the plant type. The plant type related gene obtained by the separation method is from the plant and has small influence on the environment. The separated gene is utilized to carry out rape plant type improvement molecular breeding, and the method has very important significance for cultivating new cabbage type rape varieties with improved plant types, and improving the yield and harvest index of the cabbage type rape.
The foregoing shows and describes the general principles, essential features, and advantages of the invention. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, which are described in the specification and illustrated only to illustrate the principle of the present invention, but that various changes and modifications may be made therein without departing from the spirit and scope of the present invention, which fall within the scope of the invention as claimed. The scope of the invention is defined by the appended claims and equivalents thereof.
Sequence listing
<110> Yangzhou university
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Claims (1)
1. The cabbage type rape plant type regulation protein formed by an amino acid sequence shown in SEQ ID NO. 2 or the gene which is shown in SEQ ID NO. 1 and used for coding the protein or an expression cassette, a recombinant vector, a recombinant microorganism or a transgenic cell line which contain the gene are applied to the regulation of the number of main stems of the cabbage type rape, and the regulation of the number of the main stems of the cabbage type rape is to increase the number of the main stems of the cabbage type rape.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201910092178.6A CN109810182B (en) | 2019-01-30 | 2019-01-30 | BnLAX1.c gene, protein and application thereof in controlling cabbage type rape plant type |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201910092178.6A CN109810182B (en) | 2019-01-30 | 2019-01-30 | BnLAX1.c gene, protein and application thereof in controlling cabbage type rape plant type |
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| Publication Number | Publication Date |
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| CN109810182A CN109810182A (en) | 2019-05-28 |
| CN109810182B true CN109810182B (en) | 2022-06-10 |
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| WO2007103956A2 (en) * | 2006-03-07 | 2007-09-13 | Pioneer Hi-Bred International, Inc. | Compositions and methods for increasing plant tolerance to high population density |
| CN107988252A (en) * | 2018-01-22 | 2018-05-04 | 中国科学院遗传与发育生物学研究所 | Application of the bHLH73 albumen in regulation and control plant strain senior middle school |
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| CN107988252A (en) * | 2018-01-22 | 2018-05-04 | 中国科学院遗传与发育生物学研究所 | Application of the bHLH73 albumen in regulation and control plant strain senior middle school |
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| Title |
|---|
| Overexpression of the JAZ factors with mutated jas domains causes pleiotropic defects in rice spikelet development;Yutaro Hori;《Plant Signaling & Behavior》;20141222;第9卷(第10期);e970414 * |
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