Application and method of arabidopsis thaliana gene AT1G12064 in arabidopsis thaliana breeding

Abstract

The invention discloses application of an arabidopsis thaliana gene AT1G12064 in arabidopsis thaliana breeding and a method, and belongs to the field of gene engineering. Through verification, the gene negatively regulates and controls the size and weight of arabidopsis seeds and the number of grains per pod, and positively regulates and controls the pod length, the flowering period and the growth period. A mutant of the gene is knocked out, the seed length, width, area, grain weight, grain number per pod and single plant yield are remarkably increased, the flowering period and the growth period are shortened, and early maturing and high yield are achieved. New gene resources and technical support are provided for early-maturing and high-yield breeding of cruciferous crops such as arabidopsis thaliana, oilseed rape and Chinese cabbage, and the application prospect is wide.

Description

Technical Field

[0001] This invention relates to the field of genetic engineering technology, specifically to an Arabidopsis gene. AT1G12064 Application and methods in Arabidopsis thaliana breeding. Background Technology

[0002] Arabidopsis thaliana is an annual herbaceous plant belonging to the Brassicaceae family. Its small genome, short growth cycle, high fruit production, and strong vitality make it a model plant for dicotyledonous plant research and widely used as genetic analysis material for various crops. Therefore, research on Arabidopsis thaliana can provide a research foundation and reference for other crop research and molecular breeding.

[0003] Important yield traits in Arabidopsis thaliana are typical quantitative traits, including the number of effective siliques, the number of seeds per silique, and the thousand-seed weight. The number of effective siliques is easily affected by environmental and cultivation conditions, making its genetic analysis difficult. The thousand-seed weight and the number of seeds per silique, as yield components of Arabidopsis thaliana, are less affected by environmental and cultivation conditions and are genetically more stable, making them more suitable for genetic improvement. The growth period is a crucial agronomic trait determining the planting region and season of a variety. Breeding early-maturing, high-yielding new varieties has always been a major focus of crop genetics and breeding research (Fang et al., 2019; https: / / doi.org / 10.1073 / pnas.1815030116). Overcoming latitudinal limitations in crop cultivation and reducing the constraints of regional environment and seasonal climate both highly depend on the early maturity of crops. However, early maturity of crops is negatively correlated with traits such as yield and resistance, making it difficult to simultaneously aggregate desirable traits such as early maturity, disease resistance, and yield (Zhao et al., 2023; https: / / doi.org / 10.1016 / j.tplants.2023.04.011.), which is a challenge currently facing crop breeding technology.

[0004] Currently, there is limited cloning and research on genes that control plant yield and growth period, and there is an urgent need to conduct functional research on genes that control early maturity and high yield.

[0005] It should be noted that the information disclosed in the background section above is only used to enhance the understanding of the background of this disclosure, and therefore may include information that does not constitute prior art known to those skilled in the art. Summary of the Invention

[0006] The purpose of this invention is to provide an Arabidopsis gene. AT1G12064 Its application and methods in Arabidopsis thaliana breeding provide new technical support for plant optimization breeding, and have significant applications, especially in Arabidopsis thaliana.

[0007] To achieve the above objectives, the present invention provides an Arabidopsis gene. AT1G12064 In regulating Arabidopsis seed size, knocking out this gene increases seed length, width, and area; the gene involved... AT1G12064 The nucleotide sequence is shown in SEQ ID NO.1.

[0008] This invention also provides Arabidopsis thaliana genes. AT1G12064 In regulating Arabidopsis yield, knocking out this gene can increase the thousand-seed weight, number of seeds per pod, and economic yield per plant; the gene involved... AT1G12064 The nucleotide sequence is shown in SEQ ID NO.1.

[0009] This invention also provides an Arabidopsis gene A T1G12064 Its application in regulating the growth period of Arabidopsis thaliana shows that knocking out this gene can shorten the flowering and growth periods; the gene involved... AT1G12064 The nucleotide sequence is shown in SEQ ID NO.1.

[0010] This invention also provides Arabidopsis thaliana genes. AT1G12064 Application in the creation of high-yielding and / or early-maturing Arabidopsis thaliana lines, including the genes involved. AT1G12064 The nucleotide sequence is shown in SEQ ID NO.1.

[0011] This invention also provides a method for cultivating early-maturing, high-yielding Arabidopsis thaliana, using gene loss-of-function technology. AT1G12064 Genes were used to obtain stable genetic lines with increased thousand-seed weight and earlier flowering time by 3-5 days; among them, genes... AT1G12064 The nucleotide sequence is shown in SEQ ID NO.1.

[0012] Preferably, the gene loss-of-function technology in the above method is selected from any one of CRISPR / Cas9 gene editing, T-DNA insertion mutation, or RNA interference.

[0013] This invention also provides Arabidopsis thaliana genes. AT1G12064 Its application in breeding early-maturing and high-yielding cruciferous crops, including Arabidopsis thaliana, rapeseed, Chinese cabbage, mustard greens, cauliflower, and radish. The genes involved... AT1G12064 The nucleotide sequence is shown in SEQ ID NO.1.

[0014] The present invention also provides a kit for breeding cruciferous crops, comprising a gene knockout component. AT1G12064 Primers, vectors, or mutant materials can be used, especially for cruciferous crops such as Arabidopsis thaliana, rapeseed, Chinese cabbage, mustard greens, cauliflower, and radish.

[0015] The present invention has the following advantages: This invention discloses Arabidopsis thaliana genes for the first time. AT1G12064 Uses. Through the analysis of wild-type Arabidopsis thaliana, AT1G12064 Phenotypic examination of gene knockout mutants and overexpression lines revealed that, compared with the wild-type control, the gene knockout mutants showed significantly increased seed width, length, area, seed weight, number of seeds per silique, and yield per plant, while silique length, flowering period, and growth period were shortened; the overexpression lines exhibited the opposite phenotype to the mutants. These results indicate that... AT1G12064 This gene negatively regulates seed size, weight, and number of seeds per silique in Arabidopsis thaliana; and positively regulates silique length, flowering period, and growth period. Knocking out this gene increases seed weight and number of seeds per silique in Arabidopsis thaliana, shortens flowering time and growth period, thus achieving early maturity and high yield. This invention is of significant value for research on early maturity and high yield in other important economic crops such as rapeseed, cabbage, mustard greens, cauliflower, and radish. Attached Figure Description

[0016] Figure 1 Arabidopsis thaliana constructed in this invention AT1G12064 In wild-type (Col-0), mutant (SALK_096397C), and overexpression lines (OE-1, OE-2) AT1G12064 Gene expression levels.

[0017] Figure 2 Arabidopsis thaliana constructed in this invention AT1G12064 Statistical results of seed length, width and area phenotypes in wild-type (Col-0), mutant (SALK_096397C) and overexpression lines (OE-1, OE-2).

[0018] Figure 3 Arabidopsis thaliana constructed in this invention AT1G12064 Phenotypic results of thousand-seed weight, number of seeds per silique, number of siliques per plant, silique length, biomass per plant, and economic yield per plant in wild-type (Col-0), mutant (SALK_096397C), and overexpression lines (OE-1, OE-2).

[0019] Figure 4 Arabidopsis thaliana constructed in this invention AT1G12064 Statistical results of flowering time and growth period of wild-type (Col-0), mutant (SALK_096397C) and overexpression lines (OE-1, OE-2). Detailed Implementation

[0020] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0021] Note: Unless otherwise specified, the experimental methods in the following examples are conventional methods, performed according to the techniques or conditions described in the literature in this field or according to the product instructions. Unless otherwise specified, the materials and reagents used in the following examples are commercially available.

[0022] The Arabidopsis thaliana culture involved in this technical solution is as follows: Arabidopsis thaliana seeds are placed in 1.5 mL centrifuge tubes and disinfected with 70% ethanol and anhydrous ethanol for 10 min each. The seeds are then spread on sterilized filter paper, and after the ethanol evaporates, they are evenly sprinkled onto 1 / 2 MS medium. Vernalization is then carried out at 4 °C for 48 hours, and finally, the seeds are placed in a light incubator (22 °C, 16 h daytime and 8 h nighttime) for 7–10 days. Well-grown seedlings are then transplanted into nutrient soil for further cultivation.

[0023] Example 1 Arabidopsis thaliana AT1G12064 Obtaining and identifying gene mutants Obtained from the Arabidopsis TAIR (TAIR - Home Page (arabidopsis.org)) database AT1G12064 The T-DNA insertion mutant of the gene (accession ID: SALK_096397C) has the nucleotide and amino acid sequences shown in SEQ ID NO.1 and SEQ ID NO.2, respectively.

[0024] AT1G12064 Gene (SEQ ID NO.1): ATGAAGAAGACCTACAAGCTTCAAAGCCTCTTCTCTTCTCTCATCTTCCTCATAATCTTGCTATTGATTTCGAGAGCCGCGGCCGTCGGCTCTGGCGGTGTCTGCCGGCATCCACCATCACAAAACAGCTGCAAGACATGCATGGCGGAGCAAATGAAATACGAC TGCCCTAAGTGCGTGCCGGTGCTCCGATGCATGGCTCGTTGTCTTTGGGGCGGTGTTACTCAGAGGAAGTGCACCACCACGTGTCGATGCGACACCGCGGCCAAGCCGTCGTTGCTGGAGTGTAAACGCTGCGTTTCTAGGTGTAAGTGTAGCTGTGCGGCTTAG.

[0025] AT1G12064 The gene-encoded protein (SEQ ID NO.2): MKKTYKLQSLFSSLIFLIILLLISRAAAVGSGGVCRHPPSQNSCKTCMAEQMKYDCPKCVPVLRCMARCLWGGVTQRKCTTTCRCDTAAKPSLLECKRCVSRCKCSCAA.

[0026] Extraction of genomic DNA from the mutant Arabidopsis thaliana: After Arabidopsis thaliana had grown for four weeks, a fingernail-sized piece of rosette leaf tissue was taken and placed into a 2 mL centrifuge tube containing steel balls. The Edwards method was used to crudely extract DNA from the Arabidopsis leaf for identification. The specific steps are as follows:

[0027] (1) Take one tender leaf into a 2.0 mL centrifuge tube, add two 3.5 mm steel balls, then add 500 μL of the pre-prepared Edwards extract, and break the tissue into pieces at room temperature using a high-speed sampler (sample parameters: 60 Hz, 30 s, 2 times). (2) After sampling, centrifuge at 4 °C and 12,000 rpm for 10 min; (3) Pipette 100 μL of supernatant into a 96-well PCR plate. (4) Pipette 100 μL of supernatant into a new PCR plate, add an equal volume of pre-cooled isopropanol to each well, and place in a -20 °C freezer for 30 min to precipitate DNA; centrifuge at 4000 r / min for 10 min. (5) Discard the supernatant, retain the precipitate, and air dry at room temperature; (6) Add 50 μL of sterile water to dissolve the DNA at room temperature and store at -20 °C for later use.

[0028] PCR identification: The three-primer method was used for identification. The mutant number was entered into SIGNAL (http: / / signal.salk.edu / tdnaprimers.2.html) to obtain the primer sequences for identifying the SALK_096397C mutant, as follows: LP (SEQ ID NO.3): 5'-TGGTTTTCCTTGGAGAGTTC-3' RP (SEQ ID NO.4): 5'-TGACCGGAACAAAATACTTGTG-3' LBb1.3 (SEQ ID NO.5): 5'-ATTTTGCCGATTTCGGAAC-3' PCR system: 10 μL PCR mix, 0.5 μL primer 1, 0.5 μL primer 2, 0.5 μL primer 3, 2 μL DNA template, 6.5 μL ddH2O.

[0029] PCR program: 94 °C pre-denaturation for 5 min, 94 °C denaturation for 30 s, 58 °C annealing for 30 s, 72 °C extension for 1 min, 35 cycles; 72 °C final extension for 5 min.

[0030] Agarose gel electrophoresis: PCR products were electrophoresed in a 1% agarose gel for 15 min and stained with ethidium bromide (EB). In the electrophoresis results, the homozygous wild-type amplified one band of approximately 1200 bp, the heterozygous wild-type amplified two bands of 1200 bp and 500 bp, and the homozygous mutant amplified one band of approximately 500 bp. The identified homozygous mutant is the one described above. AT1G12064 Gene mutant. That is, the two primers used in this invention span the T-DNA insertion site (LP and RP), and the other primer is on the TDNA (LBb1.3). If there is TDNA insertion at the target gene, a combined sequence of gene + TDNA of about 500 bp will be amplified; if there is no insertion, only about 1200 bp on the gene will be amplified.

[0031] Example 2 Arabidopsis thaliana AT1G12064 Creation of gene overexpression lines Since this gene lacks introns, Arabidopsis gDNA was used as a template to employ genes containing... Eco RI and Xba Primers with homologous arms added to the I restriction site ( AT1G12064 -qPCR-F, AT1G12064-qPCR-R (specific sequence as follows) amplifies the coding region sequence of the target gene and recovers the target fragment.

[0032] The vector pBin3GlyRed was used Eco RI (Takara) and Xba Double digestion with I (Takara): Plasmid 2 μg, 10× QuickCut Buffer 5 μL, QuickCut Restriction Eco RI 1 μL, QuickCutRestriction Xba I 1 μL, Nuclease-Free Water To total volume 50 μL.

[0033] The enzyme digestion products were electrophoresed on a 1% agarose gel for 30 min, stained with ethidium bromide (EB), and then excised from the gel and recovered. The DNA was purified using a Novizan kit, following the instructions in the product manual.

[0034] The purified gene PCR product was homologously recombinated with the vector pBinGlyRed (Novizan): 5×CE II Buffer 2 µL, Linear vector DNA 0.02×kb size of vector ng, Insert DNA 0.04×kb size of insert fragment ng, Exnase® II 1 µL, Nuclease-Free Water To total volume 10 μL. pBinGlyRed was obtained. AT1G12064 After cloning the vector, plasmids were extracted using the Novizan kit, following the instructions in the product manual.

[0035] Extract pBinGlyRed- AT1G12064 The vector plasmid was used to transform Arabidopsis thaliana using the Agrobacterium-mediated flower-dip transformation method. The specific procedures are as follows:

[0036] After transforming the positive clone plasmid into Agrobacterium competent cells GV3101, the plasmid was spread on LB agar plates and incubated upside down at 28°C for 1-2 days. Single clones were picked for expansion culture and identification by colony PCR. After the identification result was correct, the culture was expanded and placed in a shaker at 28°C for 200 rpm for 1-2 days. The bacterial culture was centrifuged at 4000 rpm for 15 min using a high-speed refrigerated centrifuge. The bacterial cells were collected and resuspended in staining buffer (5% sucrose solution with an appropriate amount of surfactant Silwet-L77). The OD600 was adjusted to 0.8-1.0 using a spectrophotometer.

[0037] Infect Arabidopsis inflorescences with the prepared inoculation solution for 30-45 seconds. After inoculation, place the Arabidopsis horizontally in a culture frame, spray a small amount of water on the surface, seal with plastic wrap and a black plastic bag to protect from light, and leave at room temperature for 24 hours. Then remove and place in a light incubator for normal cultivation. After 7-10 days of cultivation, perform a second inoculation. Seeds harvested after Arabidopsis maturity can be screened using green excitation light; seeds showing red light under a red filter are transgenic positive Arabidopsis.

[0038] Example 3 Arabidopsis thaliana AT1G12064 Gene expression level detection right AT1G12064 Gene mutants, wild-type Arabidopsis thaliana, AT1G12064 The following procedures were performed on the gene overexpression lines: (1) Extraction of RNA from Arabidopsis thaliana: 0.2 g of leaf tissue and 1 g of siliques 5, 10 and 15 days after flowering were ground into powder in liquid nitrogen and RNA was extracted using the SteadyPure Plant RNA Extraction Kit (Aikerui). The method was performed according to the product instructions.

[0039] (2) First-strand cDNA synthesis: First-strand cDNA was synthesized using a kit from Mona Biotechnology Co., Ltd. The method was performed according to the product instructions.

[0040] (3) Real-time PCR amplification: Refer to the operation manual of MonAmp™ ChemoHS qPCR Mix and run it using BIO-RAD real-time fluorescence quantitative instrument. Atactin8 For the internal reference gene (sequence below), each template and each primer pair were used in three replicate experiments.

[0041] Primer Atactin8-F (SEQ ID NO.6): GCAGACCGTATGAGCAAAG; Primer Atactin8-R (SEQ ID NO.7): GGAAGCAAGGATAGAACCAC; Primers AT1G12064 -qPCR-F (SEQ ID NO.8): CTCTCATCTTCCTCATAATCTTGCT; Primers AT1G12064 -qPCR-R (SEQ ID NO.9): GTATTTCATTTGCTCCGCCAT.

[0042] The results of fluorescence quantification are as follows Figure 1As shown, in the mutant (SALK_096397C) AT1G12064 The expression levels of the gene in leaves, 5-day siliques, 10-day siliques, and 15-day siliques were all significantly lower than those in the wild type (Col-0), while the expression levels in overexpression lines (OE-1, OE-2) were significantly lower. AT1G12064 The gene expression levels in leaves, 5-day, 10-day, and 15-day siliques were all significantly higher than in the wild type (Col-0). Furthermore, as the siliques developed, the expression levels in the overexpressing lines (OE-1, OE-2) increased. AT1G12064 Gene expression levels gradually increase.

[0043] Example 4: Statistical analysis of Arabidopsis thaliana seed size phenotype Wild-type (Col-0), mutant (SALK_096397C), and overexpression lines (OE-1, OE-2) were planted in the same culture frame and cultured in an artificial climate incubator. At the Arabidopsis silique maturity stage, 10-12 siliques from the main inflorescence of each plant were harvested, dried, and their pericarps removed. The siliques were then laid flat on A4 paper, numbered, and photographed. Seed images were collected using a stereomicroscope, and the length, width, and area of ​​the seeds were measured using ImageJ image processing software. Statistical results showed that, compared with the wild-type (Col-0), [the following was observed]: deletion [of certain characteristics]. AT1G12064 Genes that cause increased seed length and width, overexpression AT1G12064 Genetic factors can reduce seed length and width (see below) Figure 2 (A and B in the original text). Specifically, missing... AT1G12064 The gene increased seed length and width by 6.82% and 8.13%, respectively, and its overexpression... AT1G12064 The gene reduced seed length and width by 8.45%, 4.58%, and 9.05%, 13.24%, respectively (see [link to gene]). Figure 2 (C and D in the original text). Accordingly, missing. AT1G12064 The gene increased seed area by 15.43%, and overexpression... AT1G12064 The gene reduced the seed area by 18.86% and 19.46%.

[0044] Example 5: Statistical analysis of Arabidopsis thaliana plant yield-related phenotypic characteristics At the maturity stage of Arabidopsis thaliana, 10-15 healthy, disease-free, and undamaged individual plants were selected from each material, and their biological yield and number of siliques per plant were investigated. 10-12 siliques were taken from the main inflorescence of each individual plant, their length was measured, and the number of seeds per silique was counted. For seed weight determination, well-developed siliques from the middle of the main inflorescence were harvested and naturally dried in a 37℃ constant temperature incubator. The collected seeds from each plant were evenly spread on white paper, and the number of seeds was read using ImageJ software, while the seed weight was measured using an electronic precision balance. The economic yield per plant was calculated as the total weight of all seeds harvested from each individual plant.

[0045] The results are as follows Figure 3 As shown, compared with the wild type (Col-0), the mutant (SALK_096397C) showed significantly increased thousand-seed weight, number of seeds per silique, number of siliques per plant, and economic yield per plant, while the overexpression lines (OE-1, OE-2) showed significantly decreased values ​​for all of the above indicators (see...). Figure 3 In contrast, the silique length of the mutant (SALK_096397C) was significantly lower than that of the wild type (Col-0), while the silique length of the overexpression lines (OE-1, OE-2) was significantly increased (see AC, F). Figure 3 In addition, the biomass of the mutant (SALK_096397C) and the overexpression lines (OE-1, OE-2) was not significantly different from that of the wild type (Col-0) (see D). Figure 3 (E).

[0046] The specific data are as follows: the thousand-seed weight, number of seeds per silique, number of siliques per plant, and economic yield per plant for the wild type (Col-0) were 21.30±0.56 mg, 47.52±2.94 seeds, 189.10±57.23 seeds, and 47.39±9.64 mg, respectively. The corresponding values ​​for the mutant (SALK_096397C) were 22.69±0.66 mg, 52.38±2.72 seeds, 238.70±28.75 seeds, and 59.20±14.39 mg, respectively, which were significantly higher than those for the wild type (Col-0) by 6.51%, 10.23%, 26.23%, and 24.92%. The mean thousand-seed weight, number of seeds per silique, number of siliques per plant, and economic yield per plant of the overexpression lines (OE-1 and OE-2) were 19.75±0.70 mg and 19.40±1.57 mg, 46.13±1.53 seeds and 44.69±2.92 seeds, 13.89±23.83 seeds and 140.40±25.07 seeds, and 37.44±6.75 mg and 32.98±11.45 mg, respectively, which were significantly lower than those of the wild type (Col-0) by 7.29% and 8.95%, 2.92% and 5.95%, 26.55% and 25.75%, and 20.98% and 30.41%, respectively.

[0047] Regarding silique length, the wild type (Col-0) was 1.23±0.03 cm; the mutant (SALK_096397C) was 1.13±0.06 cm, significantly lower than the wild type (Col-0) by 8.47%; the overexpression lines (OE-1 and OE-2) were 1.27±0.03 cm and 1.27±0.03 cm, respectively, significantly higher than the wild type (Col-0) by 3.32% and 3.13%. There was no significant difference in biomass per plant among the wild type (52.64±65.65 mg), mutant (252.64±65.65 mg), and overexpression lines OE-1 (249.04±55.49 mg) and OE-2 (242.27±43.91 mg).

[0048] Example 6: Phenotypic Statistics and Analysis of Arabidopsis thaliana Plants During Flowering and Growth Stages Flowering time was defined as the number of days from sowing date until at least 25% of the plants in each material had at least three open flowers. The growth period was defined as the total number of days from sowing to harvest. Results are shown below. Figure 4 As shown in A and B, the flowering time and growth period of the mutant (SALK_096397C) were significantly shorter than those of the wild type (Col-0), while the flowering time and growth period of the overexpression lines (OE-1, OE-2) were significantly longer than those of the wild type (Col-0).

[0049] The specific data are as follows: the flowering period and growth period of the wild type (Col-0) were 44.64±2.27 days and 66.55±2.12 days, respectively. The flowering period and growth period of the mutant (SALK_096397C) were 40.33±2.84 days and 60.92±3.38 days, respectively, significantly shorter than the wild type by 9.64% and 8.46%. The flowering period of the overexpression line OE-1 was 51.23±2.75 days, and the growth period was 70.40±1.82 days; the flowering period of OE-2 was 53.87±2.58 days, and the growth period was 69.53±3.70 days. Compared with the wild type, the flowering period of OE-1 was significantly delayed by 14.77%, and that of OE-2 by 20.68%; in terms of growth period, OE-1 was significantly prolonged by 5.79%, and that of OE-2 by 4.49%.

[0050] Although the present invention has been described in detail through the preferred embodiments above, it should be understood that the above description should not be considered as a limitation of the present invention. Various modifications and substitutions to the present invention will be apparent to those skilled in the art after reading the above description. Therefore, the scope of protection of the present invention should be defined by the appended claims.

Claims

1. Arabidopsis gene AT1G12064 use in modulating seed size in Arabidopsis, characterized in that, Knocking out the gene can increase the length, width and area of the seed; wherein the nucleotide sequence of the gene is shown as SEQ ID NO.

1. AT1G12064 the nucleotide sequence of the gene is shown as SEQ ID NO.

1.

2. Arabidopsis gene AT1G12064 use in modulating yield in Arabidopsis, characterized in that, Knocking out the gene can increase the thousand-grain weight, the number of grains per silique, and the economic yield per plant; wherein the nucleotide sequence of the gene is shown as SEQ ID NO.

1. AT1G12064 the nucleotide sequence of the gene is shown as SEQ ID NO.

1.

3. Arabidopsis gene A T1G12064 In the regulation of the growth period of Arabidopsis thaliana, characterized in that, Knocking out the gene can shorten the flowering period and the growth period; wherein the nucleotide sequence of the gene is shown as SEQ ID NO.

1. AT1G12064 Knocking out the gene can shorten the flowering period and the growth period; wherein the nucleotide sequence of the gene is shown as SEQ ID NO.

1.

4. Arabidopsis gene AT1G12064 In the creation of high yielding or / and early maturing Arabidopsis lines, wherein, The nucleotide sequence of the gene AT1G12064 is shown as SEQ ID NO.

1.

5. A method for breeding early-maturing, high-yielding Arabidopsis thaliana, characterized by, Adopting gene function loss technology to treat AT1G12064 Gene, obtain stable genetic strain of 1000-grain weight increase and flowering time 3-5 days in advance; wherein, the nucleotide sequence of the gene AT1G12064 As shown in SEQ ID NO.

1.

6. The method of claim 5, wherein, The gene loss-of-function technique is selected from any one of CRISPR / Cas9 gene editing, T-DNA insertion mutation or RNA interference.

7. Arabidopsis gene AT1G12064 application in breeding of early maturity and high yield of Brassica crops; wherein, The nucleotide sequence of the gene AT1G12064 is shown as SEQ ID NO.

1.

8. Use according to claim 7, characterized in that, The Brassicaceae crop comprises Arabidopsis thaliana, Brassica napus, Brassica rapa, Brassica juncea, Brassica oleracea, Raphanus sativus.

9. A kit for breeding plants of the family Brassicaceae, characterized in that, The kit comprises primers, vectors or mutant materials for knocking out a gene AT1G12064 ; wherein the nucleotide sequence of the gene AT1G12064 is shown as SEQ ID NO.

1.

10. The kit of claim 9, wherein The Brassicaceae crop comprises Arabidopsis thaliana, Brassica napus, Brassica rapa, Brassica juncea, Brassica oleracea, Raphanus sativus.

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