Gene and method for regulating branch angle of brassica napus and application thereof

Abstract

The application provides genes and regulation methods and applications for regulating the branching angle of oilseed rape, and belongs to the technical field of functional genes.The application provides genes BnTAC1-1 and / or BnTAC1-2 for regulating the branching angle of oilseed rape, and the application finds that the average branching angle of double-gene-edited oilseed rape is smaller than that of single-gene-edited oilseed rape, and the average branching angle of single-gene-edited oilseed rape is smaller than that of wild-type oilseed rape by creating gene editing mutants.It is proved that the genes BnTAC1-1 and BnTAC1-2 can jointly control the branching angle of oilseed rape, and have additive effects, so as to regulate the compactness of the plant type of Brassica napus, and will not affect the flowering period, the number of branches, the number of silique and the number of silique grains and other agronomic characters of oilseed rape, and can be applied to the improvement of oilseed rape germplasm, and the creation of oilseed rape suitable for high-density planting, and the improvement of the yield of oilseed rape per unit area.

Description

TECHNICAL FIELD

[0001] The application belongs to the field of functional gene technology, and particularly relates to a gene for regulating the branching angle of Brassica napus and a regulation method and application. BACKGROUND

[0002] Brassica napus is an important oil crop and an important source of plant protein for feeding, and has a large planting area in the world. In recent years, the planting area of Brassica napus in China is nearly 7 million mu, accounting for about one third of the total global planting area. Although the planting area of Brassica napus in China has been increasing year by year, there is a problem of idle farmland in the Yangtze River Basin in winter due to income issues in the traditional Brassica napus production area. Therefore, it is currently urgent to increase the yield per unit area of Brassica napus to improve the planting efficiency of Brassica napus, so as to further increase the planting area of Brassica napus.

[0003] Ultra-high density and ultra-high yield are the main direction of Brassica napus yield increase, and Brassica napus breeders generally believe that compact Brassica napus is the "ideal plant type" to achieve high yield of Brassica napus. Brassica napus has a large plant height, long branches and many angles, forming a unique angle layer to store yield. After the flowering period of Brassica napus, the dense angle layer will affect the photosynthesis of the leaves and the air exchange of the whole population, and the weakened photosynthesis and insufficient carbon dioxide concentration will affect the yield of Brassica napus. The size of the branching angle directly affects the degree of intersection of the angle layer in the Brassica napus field, and a reasonable branching angle can reduce the intersection of branches between different plants, thereby reducing the intersection degree of the angle layer, increasing the ventilation performance and light transmission performance of the whole population, and being beneficial to yield increase. In addition, a properly compact plant type can be reasonably planted at high density, which is also beneficial to yield increase; and the reduction of the vertical and horizontal intersection of branches between plants can reduce the damage to plants caused by personnel activities during field management, and is also beneficial to reducing the harvest loss caused by the bursting of angles due to the winding of branches during mechanical harvesting, which are all beneficial to ensuring the harvest yield. In summary, it has important practical significance to tap the Brassica napus branching angle gene for cultivating Brassica napus varieties with high density and mechanical harvesting, increasing the yield of Brassica napus, and ensuring the safety of edible oil in China. SUMMARY

[0004] The application provides a gene for regulating the branching angle of Brassica napus and a regulation method and application, and the gene BnTAC1-1 and / or BnTAC1-2 can regulate the branching angle of Brassica napus, thereby helping to achieve ultra-high density and ultra-high yield of Brassica napus cultivation.

[0005] The application provides a gene for regulating the branching angle of Brassica napus, including BnTAC1-1 and / or BnTAC1-2, wherein the amino acid sequence encoded by the BnTAC1-1 includes SEQ ID No. 1 or a sequence having 85% or more homology with the sequence described in SEQ ID No. 1;

[0006] The amino acid sequence coded by the BnTAC1-2 includes SEQ ID No. 2 or a sequence homologous to SEQ ID No. 2 with more than 85%.

[0007] In one preferred mode of the present application, the genomic nucleotide sequences of BnTAC1-1 and BnTAC1-2 are as shown in SEQ ID No. 3 and SEQ ID No. 4.

[0008] In one preferred mode of the present application, the CDS sequences of BnTAC1-1 and BnTAC1-2 are as shown in SEQ ID No. 5 and SEQ ID No. 6.

[0009] The present application also provides an application of the above-mentioned genes in regulating the plant type of Brassica napus.

[0010] In one preferred mode of the present application, the regulation of the plant type of Brassica napus includes that after knocking out one gene, the branching angle of Brassica napus is reduced and the plant type of Brassica napus is compact; after knocking out two genes, the branching angle of Brassica napus is smaller and the plant type of Brassica napus is more compact.

[0011] In one preferred mode of the present application, the Brassica napus includes Brassica napus L.

[0012] The present application also provides an application of the above-mentioned genes in creating Brassica napus germplasm suitable for high-density planting.

[0013] The present application also provides a method for creating Brassica napus germplasm suitable for high-density planting, which comprises knocking out the above-mentioned genes in the genome of Brassica napus.

[0014] The present application also provides sgRNAs for knocking out the above-mentioned genes, including sgRNA1 and sgRNA2, wherein the nucleotide sequence of sgRNA1 is as shown in SEQ ID No. 7 and the nucleotide sequence of sgRNA2 is as shown in SEQ ID No. 8.

[0015] The present application also provides gene editing vectors containing the above-mentioned sgRNAs.

[0016] The present application also provides an application of the Brassica napus germplasm suitable for high-density planting obtained by the above-mentioned method in improving the yield of Brassica napus.

[0017] Beneficial effects: the present application provides two homologous genes BnTAC1-1 and BnTAC1-2 of TAC1 in oilseed rape, in the embodiments of the present application, by constructing a CRISPR / Cas9 gene editing vector and carrying out genetic transformation on the main leading variety of Zhejiang province oilseed rape Zheyou 51, finally, a gene edited oilseed rape in which both BnTAC1-1 and BnTAC1-2 are edited, and a gene edited oilseed rape in which only BnTAC1-1 is edited are obtained, and then phenotype identification is carried out, it is found that the average branch angle of wild type Zheyou 51 is 36.2 degrees, the average branch angle of single gene edited oilseed rape is 29.9 degrees, and the average branch angle of double gene edited oilseed rape is 24.4 degrees. It is proved that the genes BnTAC1-1 and BnTAC1-2 described in the present application can control the size of the branch angle of oilseed rape, and have additive effect, so as to regulate the plant type of Brassica napus, and the flowering period, the number of branches, the number of silique and the number of silique grains have no difference with the wild type, more amount of oilseed rape can be planted on unit area, and the yield per unit area is improved. BRIEF DESCRIPTION OF DRAWINGS

[0018] Figure 1 It is a result analysis diagram of double gene edited oilseed rape editing type;

[0019] Figure 2 It is a result analysis diagram of single gene edited oilseed rape editing type;

[0020] Figure 3 It is a phenotype comparison (A) of gene edited oilseed rape and wild type oilseed rape, average vertical projection area (B) of oilseed rape plant and branch angle diagram (C) of oilseed rape primary branch and main stem;

[0021] Figure 4 It is a gene knockout strategy diagram of the present application. DETAILED DESCRIPTION

[0022] The present application provides a gene for regulating the branch angle of oilseed rape, including BnTAC1-1 and / or BnTAC1-2, wherein the amino acid sequence encoded by BnTAC1-1 includes SEQ ID No. 1 or a sequence having more than 85% homology with the sequence described in SEQ ID No. 1;

[0023] The amino acid sequence encoded by BnTAC1-2 includes SEQ ID No. 2 or a sequence having more than 85% homology with the sequence described in SEQ ID No. 2.

[0024] In an embodiment of the present application, the amino acid sequence encoded by BnTAC1-1 is as shown in SEQ ID No. 1:

[0025] MYFHVVKEIDGVAKSEKKKRGEGTSEIEKNTKAIMDQVGLVDALDNWFDGVLTIGTF GFDTLKFQEEAEIDDGDERESVGLDYVVIDGSIIKNVNQESDPLISNENKVYDHHED LEALCINHFESVKTVERAVVVAAAAAETEVEPEKKRTTLAELFMEDRVKDDDTKHDK KKPKNRNLDVDGQEIKYHKQNGSKLPSKLSFAKKMIITKSKDTEDSRPIKNEHDSRP IKKVHQMIKRMLKKKIHPDMDASKASKKDGPYKPALNCEALETLYLLNVPGSSF*;

[0026] The genomic sequence of the BnTAC1-1 is shown as SEQ ID No. 3:

[0027]

[0028] The CDS sequence of the BnTAC1-1 is shown as SEQ ID No. 5:

[0029] ATGTATTTTCATGTGGTGAAAGAAATAGATGGTGTGGCGAAAAGCGAGAAAAAGAAGAGAGGTGAAGGAACAAGTGAGATAGAGAAGAACACGAAAGCTATAATGGATCAGGTTGGGTTAGTGGATGCTCTTGATAACTGGTTCGATGGAGTTCTCACCATCGGCACATTTGGTTTTGACACTTTGAAATTTCAAGAAGAAGCCGAAATAGATGATGGTGATGAACGCGAGAGTGTGGGTTTGGACTATGTGGTAATCGATGGTAGCATCATCAAGAACGTCAACCAAGAGTCGGACCCTCTTATCTCCAACGAGAATAAGGTTTATGATCATCATGAGGATTTAGAGGCATTATGCATTAATCACTTTGAGTCGGTCAAAACGGTTGAAAGGGCGGTGGTCGTGGCCGCGGCCGCGGCGGAGACCGAGGTGGAGCCGGAGAAGAAGAGGACAACACTAGCCGAGCTTTTCATGGAAGACCGAGTAAAAGACGATGATACGAAGCATGACAAGAAGAAACCAAAGAACCGTAATCTTGATGTTGATGGCCAGGAGATTAAATATCATAAACAAAACGGGTCAAAGCTGCCTAGTAAGTTATCGTTTGCTAAGAAGATGATCATTACTAAGTCCAAAGATACAGAAGACTCGCGTCCAATCAAGAACGAGCATGACTCGCGTCCAATCAAGAAAGTGCATCAGATGATAAAGAGGATGTTAAAGAAGAAGATCCATCCAGATATGGATGCGAGTAAGGCGTCTAAAAAGGACGGTCCATACAAGCCAGCCCTGAATTGTGAAGCTCTCGAGACACTTTATCTTCTTAATGTTCCAGGTTCCTCTTTTTGA.

[0030] The application also provides the amplification primer sequence of BnTAC1-1, including the upstream primer BnaTAC1-1-F with the nucleotide sequence as shown in SEQ ID No. 9 and the downstream primer BnaTAC1-1-R with the nucleotide sequence as shown in SEQ ID No. 10:

[0031] BnaTAC1-1-F: ACTGGGTGCAGAAGAAGCTG;

[0032] BnaTAC1-1-R: CGACGTCGTTCTGAGACTTC.

[0033] In one embodiment of the application, the protein sequence encoded by BnTAC1-2 is shown as SEQ ID No. 2: MSSEKHVYFHVVKEIDGVAKSEKKKRGEGTSEIEKNTKAILDQVGLVDALDNWFDGVLTIGTFGFDTLKFQEEAEIDDGDECESVGLDYVVVDGSIIKNVNQESDPLISNENKVYDHHEDLEALCINHFESVKTVERPVIVAAAEAEVEPEKKRTTLAELFLEDRVKDDDTKHDKKKPKNRNLDVDGQEVKYHKQNGSKLSSKFSFAKKMIISKPKDKEDSRPIKKEHDSRPIKKVHQMIKRMLKKKIHPDMDATKASKKDGPYKPALKCEALETLYLLNVPGSSF*;

[0034] The genomic sequence of BnTAC1-2 is shown as SEQ ID No. 4:

[0035]

[0036] The CDS sequence of which is shown as SEQ ID No. 6:

[0037] ATGTCATCAGAGAAACATGTGTATTTTCATGTGGTGAAAGAAATAGATGGTGTGGCGAAAAGCGAGAAAAAGAAGAGAGGTGAAGGAACAAGTGAGATAGAGAAGAACACGAAAGCTATACTGGATCAGGTTGGGTTAGTGGATGCTCTTGATAACTGGTTCGATGGAGTTCTCACCATCGGCACATTCGGTTTTGACACTTTGAAATTTCAAGAAGAAGCCGAAATAGATGATGGTGATGAATGCGAGAGTGTGGGTTTGGACTATGTCGTAGTCGATGGTAGCATCATCAAGAACGTCAACCAAGAGTCGGACCCTCTTATCTCCAATGAGAATAAGGTTTATGATCATCATGAGGATTTAGAGGCGTTATGCATTAATCACTTTGAGTCGGTCAAAACGGTTGAAAGGCCGGTGATCGTGGCCGCGGCGGAGGCCGAGGTGGAGCCGGAGAAGAAGAGGACAACACTAGCCGAGCTTTTCCTGGAAGACCGAGTAAAAGACGATGATACGAAGCATGACAAGAAGAAACCAAAGAACCGTAATCTTGATGTTGATGGCCAGGAGGTTAAATATCATAAACAAAACGGGTCAAAGCTGTCTAGTAAGTTCTCCTTTGCTAAGAAGATGATCATTTCAAAGCCCAAAGATAAAGAAGACTCGCGTCCAATCAAGAAAGAGCATGACTCGCGTCCAATCAAGAAAGTGCATCAGATGATAAAGAGGATGCTAAAGAAGAAGATTCATCCAGATATGGATGCGACTAAGGCTTCTAAAAAGGACGGTCCATACAAGCCAGCCCTGAAATGTGAAGCTCTCGAGACACTTTATCTTCTTAATGTTCCAGGTTCGTCTTTTTGA.

[0038] The application also provides an amplification primer sequence of BnTAC1-2, including an upstream primer BnaTAC1-2-F with a nucleotide sequence as shown in SEQ ID No. 11 and a downstream primer BnaTAC1-2-R with a nucleotide sequence as shown in SEQ ID No. 12:

[0039] BnaTAC1-2-F: ACTGATTGGTATTGTCTCCC;

[0040] BnaTAC1-2-R: CACGTTGTCACATAATAATAAAAT.

[0041] The application also provides an application of the above gene in regulating the plant type of rapeseed.

[0042] In a preferred mode of the application, the regulation of the plant type of rapeseed includes that after the knockout of the gene, the branching angle of rapeseed is reduced, and the plant type of rapeseed is compact. In an embodiment of the application, by constructing a CRISPR / Cas9 gene editing vector and performing genetic transformation on Zheyou 51, a gene edited rapeseed in which both BnTAC1-1 and BnTAC1-2 are edited and a gene edited rapeseed in which only BnTAC1-1 is edited are finally obtained. In the embodiment of the application, it is found through phenotype identification that the average branching angle of wild type Zheyou 51 is 36.2 degrees, the average branching angle of the double gene edited rapeseed is 24.4 degrees, and the average branching angle of the single gene edited rapeseed is 29.9 degrees. In conclusion, the application proves that both the genes BnTAC1-1 and BnTAC1-2 can control the size of the branching angle of rapeseed, and have additive effects, which can achieve a better effect of controlling the branching angle of rapeseed.

[0043] The application also provides sgRNA used for knocking out the gene, including sgRNA1 and sgRNA2, wherein the nucleotide sequence of sgRNA1 is as shown in SEQ ID No. 7, and the nucleotide sequence of sgRNA2 is as shown in SEQ ID No. 8.

[0044] sgRNA1: GAGTGTGGGTTTGGACTATGTGG;

[0045] sgRNA2: AGATAAGAGGGTCCGACTCTTGG.

[0046] In the application, after overexpression of the gene, the plant type of rapeseed is loose, and the branching angle of rapeseed is increased.

[0047] In one preferred mode of the present application, the oilseed rape includes Brassica napus, such as Zheyou 51 verified in the examples, which is an oilseed rape variety selected from Huyou 15 / Zheduan 6 by Crop and Nuclear Technology Utilization Institute of Zhejiang Academy of Agricultural Sciences, and passed the National Crop Variety Approval Committee of Zhejiang Province in 2009 with the approval number: Zheping 2009001.

[0048] The branch angle referred to in the present application refers to the included angle between the primary branch and the main stem, which is an important determinant of the plant type of the oilseed rape. According to observation and statistics, after the knockout of the gene, the number of branches, the number of pods, the number of grains per pod, the length of branches and the plant height have no difference compared with the wild type Brassica napus, but the branch angle is different.

[0049] The present application also provides a gene editing vector comprising the sgRNA.

[0050] The sgRNA1 acts on BnTAC1-1, and the sgRNA2 acts on BnTAC1-1 and BnTAC1-2 together, and a sequence (SEQ ID No. 13) containing sgRNA1 and sgRNA2 is synthesized: gagtgtgggtttggactatggttttagagctagaaatagcaagttaaaataaggctagtccgttatcaacttgaaaaagtggc accgagtcggtgcaacaaagcaccagtggtctagtggtagaatagtaccctgccacggtacagacccgggttcgattcccggctggtgcaagataagagggtccgactct, and the sequence shown in SEQ ID No. 13 is connected to the intermediate vector puc57, and then a primer with a CRISPR / cas9 vector connector (SEQ ID No. 14 and SEQ ID No. 15) is synthesized to amplify sgRNA1 and sgRNA2 which have been connected together, and the amplified fragment is connected to the CRISPR / cas9 vector, and after the successful construction of the vector, the DH5α strain is transformed and sequenced by first-generation Sanger sequencing for sequencing confirmation, and finally the agrobacterium strain which can be used for genetic transformation of oilseed rape containing the above-mentioned CRISPR / cas9 vector is obtained. The CRISPR / cas9 vector referred to in the present application is from the laboratory of Professor Chen Qijun of China Agricultural University, and has been disclosed in the article (Xing et al. A CRISPR / Cas9 toolkit for multiplex genome editing in plants. BMC Plant Biology. 2014, 14: 327).

[0051] Primer: F (SEQ ID No. 14):cagtggtctcatgcaGAGTGTGGGTTTGGACTATG;

[0052] R (SEQ ID No. 15):cagtggtctcaaaacAGAGTCGGACCCTCTATCT.

[0053] This invention also provides the application of the above-mentioned genes in creating rapeseed germplasm suitable for high-density planting.

[0054] In one embodiment of the present invention, a compact rapeseed plant can be constructed by gene knockout, increasing the planting density per unit area. For example, in this embodiment, the gene-edited plant and the wild-type plant are compared using the average vertical projection area, where the average vertical projection area s = πa. 2 'a' represents the vertical projection length of the branch onto the ground corresponding to different angles, i.e., the radius of the circle formed by the vertical projection of the plant; for example... Figure 3 In triangle C, the lengths c of the main stem and branches, as well as the branching angle A, are known. Using the sine function sinA = a / c, the vertical projection length 'a' of the branches on the ground corresponding to different angles can be calculated. Theoretically, this is the radius of the vertical projection circle of the rapeseed plant on the ground. Comparison showed no difference in branch length among the three different genotypes of rapeseed Zheyou 51. Calculations revealed that the average vertical projection area of ​​the rapeseed plant with the two gene deletions was 0.1859 m². 2 The average vertical projection area of ​​rapeseed plants with single-gene loss of function was 0.2702 m². 2 The average vertical projection area of ​​wild-type Zheyou 51 is 0.3780 m². 2 Compared to the wild-type Zheyou 51, the vertical projection area of ​​the double-gene loss-of-function rapeseed plant is reduced by more than 50%, meaning that the theoretical land area required is less than half that of the wild-type Zheyou 51, indicating a strong potential for increased yield through dense planting.

[0055] The present invention also provides a method for creating rapeseed germplasm suitable for high-density planting, which includes knocking out the above-mentioned genes in the rapeseed genome.

[0056] The present invention does not specifically limit the method for causing gene function loss. Gene function loss can be caused by using conventional techniques in the art. For example, the gene knockout method included above was used in the embodiments.

[0057] This invention also provides the application of rapeseed germplasm suitable for high-density planting obtained by the above-described creation method in improving rapeseed yield.

[0058] The oilseed rape germplasm suitable for high-density planting constructed by the method can increase the planting amount per unit area and does not affect the yield per plant, and therefore can significantly improve the yield of oilseed rape.

[0059] In order to further illustrate the present application, the gene for regulating the branch angle of oilseed rape and the regulating method and application provided by the present application are described in detail below in combination with examples, but they should not be understood as limiting the protection scope of the present application.

[0060] Example 1 Amplification of BnTAC1-1 and BnTAC1-2

[0061] 1. Synthesis of primer pairs

[0062] SEQ ID No. 9: BnaTAC1-1-FACT GGGTGCAGAAGAAGCTG;

[0063] SEQ ID No. 10: BnaTAC1-1-R CGACGTCGTTCTGAGACTTC;

[0064] SEQ ID No. 11: BnaTAC1-2-FACT GATTGGTATTGTCTCCC;

[0065] SEQ ID No. 12: BnaTAC1-2-R CACGTTGTCACATAATAATAAAAT.

[0066] 2. Amplification of gene sequences using KOD Plus high-fidelity enzyme, 50 μL amplification system: 10×PCR Buffer 5 μL, 2 mM dNTPs 5 μL, 2 mM MgSO4 2 μL, KOD plus ploymerase 1 μL, cDNA 2 μL, 10 pmol / μL upper and lower primers each 1.5 μL, and 32 μL ddH2O.

[0067] 3. The band size of the PCR product is detected by agarose gel (1%) electrophoresis, and the corresponding gel block is cut according to the band size, and the DNA gel recovery kit is used for purification and recovery of the PCR fragment, and Ex taq enzyme is used for tailing.

[0068] Since the KOD plus-enzyme is a high-fidelity enzyme, the product is a blunt end, and tailing experiment is needed, and the tailing reaction system (10 μL) is as follows: Ex Taq 4 μL and PCR product 6 μL. 72°C, 30 min.

[0069] 4, connected to the cloning vector pMDTM19-T, the ligation product was transformed into DH5a competent cells, cultured overnight at 37°C, and the next day colony PCR was performed to detect qualified colonies for Sanger sequencing.

[0070] Example 2B Regulation of the branch angle of oilseed rape by BnTAC1-1 and BnTAC1-2

[0071] In Figure 4 The sgRNA for gene editing target site was designed according to the gene position, wherein sgRNA1: GAGTGTGGGTTTGGACTATGTGG acts on BnTAC1-1, sgRNA2: AGATAAGAGGGTCCGACTCTTGG acts on BnTAC1-1 and BnTAC1-2, according to the sequences of sgRNA1 and sgRNA2, a vector sequence containing sgRNA1 and sgRNA2 was synthesized: gagtgtgggtttggactatggttttagagctagaaatagcaagttaaaataaggctagtccgttatcaacttgaaaaagtggc accgagtcggtgcaacaaagcaccagtggtctagtggtagaatagtaccctgccacggtacagacccgggttcgattccc ggctggtgcaagataagagggtccgactct, and the sequence was connected to the intermediate vector puc57, and then primers with a CRISPR / cas9 vector adapter were synthesized to amplify sgRNA1 and sgRNA2 that have been connected in series, and the amplified fragments were connected to the CRISPR / cas9 vector. After successful construction of the vector, it was transformed into DH5a strain and sequenced by first-generation Sanger sequencing for sequence confirmation, and finally a Agrobacterium strain containing the above-mentioned CRISPR / cas9 vector was obtained, which can be used for genetic transformation of oilseed rape.

[0072] Genetic transformation of Zheyou 51 was carried out by constructing a CRISPR / Cas9 gene editing vector, and finally Figure 1 the gene edited oilseed rape in which both BnTAC1-1 and BnTAC1-2 were edited, and Figure 2 the gene edited oilseed rape in which only BnTAC1-1 was edited. The results of phenotypic identification of the main stem and branches are shown in Table 1 and Figure 3 The average branch angle of wild-type Zheyou 51 was 36.2 degrees, the average branch angle of the double-gene edited oilseed rape was 24.4 degrees, and the average branch angle of the single-gene edited oilseed rape was 29.9 degrees. In summary, the two genes BnTAC1-1 and BnTAC1-2 in oilseed rape jointly control the size of the branch angle of oilseed rape, and have additive effects.

[0073] Table 1 Plant type of gene edited Brassica napus

[0074]

[0075] Although the above embodiments have been described in great detail, it should be understood that the application is not limited to the embodiments described, but only to the claims. Other embodiments can be obtained from the above embodiments without being inventive.

Claims

1. Gene knockout BnTAC1-1 Its application in the preparation of compact-plant rapeseed is characterized by... The BnTAC1-1 The genomic nucleotide sequence is shown in SEQ ID No.

3.

2. Simultaneously knock out genes BnTAC1-1 and BnTAC1-2 Its application in the preparation of compact-plant rapeseed is characterized by... The BnTAC1-1 The genomic nucleotide sequence is shown in SEQ ID No. 3; BnTAC1-2 The genomic nucleotide sequence is shown in SEQ ID No.

4.

3. The application according to claim 1 or 2, characterized in that, The rapeseed mentioned includes Brassica napus.

4. A method for creating rapeseed germplasm suitable for high-density planting, characterized in that, Including knockout in the rapeseed genome BnTAC1-1 Or knock out at the same time BnTAC1-1 and BnTAC1-2 ; The above BnTAC1-1 The genomic nucleotide sequence is shown in SEQ ID No. 3; The BnTAC1-2 The genomic nucleotide sequence is shown in SEQ ID No. 4.

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