Beta-d-xylosidase gene and its use for promoting coppice regeneration

Abstract

The application provides a Caragana korshinskii beta-D-xylosidase gene and application thereof in promoting stubble regeneration. The full-length of the Caragana korshinskii beta-D-xylosidase gene CDS is 2322 bp, contains 6 exons, encodes a polypeptide with 773 amino acids, and contains two glycosyl hydrolase domains. The plant overexpressing the beta-D-xylosidase gene has more vigorous growth ability, and the root is longer than that of the wild type. After stubble, the beta-D-xylosidase degrades polysaccharides in the root cell wall, and transports upwards, so as to provide energy for the regeneration process. The Caragana korshinskii beta-D-xylosidase gene and the above functions are discovered for the first time, so as to provide a target gene for analyzing the stubble regeneration mechanism of Caragana korshinskii and breeding high-quality Caragana korshinskii.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of plant genetic engineering, and particularly relates to a Caragana korshinskii beta-D-xylosidase gene and application thereof in promoting root growth, sugar transport, cell wall morphology change and stubble regeneration. BACKGROUND

[0002] Caragana korshinskii is a leguminous shrub widely distributed in arid and semi-arid regions of northwest China. Local farmers often harvest its branches for firewood, and its twigs are often used as forage for cattle and sheep. Caragana korshinskii can adapt to dry environmental conditions, but as it ages, its water conductivity decreases year by year, photosynthesis weakens, nutrient accumulation slows down, and the degree of lignification increases. If not intervened artificially, its stress resistance will decrease and it will easily dry up and die. After the aboveground branches of Caragana korshinskii are cut off, it usually undergoes a regeneration process, rapidly generating multiple new sprouts at the rootstock junction, improving water conductivity, and rapidly accumulating nutrients in the regenerated sprouts and leaves. After 7-9 years of regrowth, its height and biomass can be restored. Stubble cutting is an important measure for the rational use of Caragana korshinskii biological resources and the protection of the environment. The applicant's research group has previously conducted a large amount of research on the physiological mechanism of Caragana korshinskii stubble cutting, and has elucidated the change rules of water use efficiency, photosynthetic rate and transpiration rate after stubble cutting of Caragana korshinskii, but the genetic and molecular mechanisms are still unclear.

[0003] Beta-D-xylosidase belongs to the glycoside hydrolase family and plays a crucial role in the decomposition of xylan and hemicellulose. The applicant screened differential expression genes after stubble cutting of Caragana korshinskii by transcriptome sequencing technology, and verified several key genes in the stubble regeneration process by qRT-PCR, and found that beta-D-xylosidase was significantly up-regulated at the transcriptional level. This may be related to the reuse of hemicellulose by beta-D-xylosidase to provide material and energy reserves for regeneration. The present application also constructs a plant expression vector of the stubble regeneration key gene beta-D-xylosidase and transforms model plants to verify the gene function. The present application aims to analyze the molecular mechanism of beta-D-xylosidase in promoting stubble regeneration of Caragana korshinskii, and to provide a basis for further screening of excellent germplasm and playing its ecological and production functions.

[0004] Problems existing in the prior art: There is no report on Caragana korshinskii beta-D-xylosidase gene and its function in the prior art. SUMMARY

[0005] In view of the deficiencies in the prior art, the present application provides a Caragana korshinskii beta-D-xylosidase gene and application thereof in promoting root growth, sugar transport, cell wall morphology change and stubble regeneration. In order to achieve the above-mentioned purpose, the technical scheme adopted by the present application is as follows:

[0006] 1. A Caragana β-D-xylosidase gene, wherein the full-length sequence of the β-D-xylosidase gene is shown in SEQ ID NO. 1, and the transcript sequence of the β-D-xylosidase gene is shown in SEQ ID NO. 2.

[0007] 2. A method for screening Caragana stubble regeneration key genes, wherein the method comprises the following steps: (1) experimental material selection and treatment, (2) transcriptome analysis, and (3) selection of stubble regeneration key genes.

[0008] 3. A method for functional analysis of Caragana β-D-xylosidase genes, wherein the method comprises the following steps:

[0009] (1) Caragana β-D-xylosidase gene cloning and plant overexpression vector construction, (2) Caragana β-D-xylosidase gene structure analysis, (3) Caragana β-D-xylosidase gene Arabidopsis transformation, and (4) transgenic Arabidopsis phenotype and biochemical analysis.

[0010] 4. Application of overexpressing β-D-xylosidase genes to promote root growth, wherein the full-length sequence of the β-D-xylosidase gene is shown in SEQ ID NO. 1, and the transcript sequence of the β-D-xylosidase gene is shown in SEQ ID NO. 2.

[0011] 5. Application of overexpressing β-D-xylosidase genes to change cell wall morphology, wherein the full-length sequence of the β-D-xylosidase gene is shown in SEQ ID NO. 1, and the transcript sequence of the β-D-xylosidase gene is shown in SEQ ID NO. 2; and the change in cell wall morphology refers to reducing the thickness and density of the cell wall and increasing the number of pits.

[0012] 6. Application of overexpressing β-D-xylosidase genes to promote sugar transport after stubble, wherein the full-length sequence of the β-D-xylosidase gene is shown in SEQ ID NO. 1, and the transcript sequence of the β-D-xylosidase gene is shown in SEQ ID NO. 2; and the sugar transport refers to transport from bottom to top, from the root to the rootstock connection.

[0013] 7. Application of overexpressing β-D-xylosidase genes to promote stubble regeneration, wherein the full-length sequence of the β-D-xylosidase gene is shown in SEQ ID NO. 1, and the transcript sequence of the β-D-xylosidase gene is shown in SEQ ID NO. 2.

[0014] 8. A vector comprising a Caragana β-D-xylosidase gene CDS sequence, wherein the full-length sequence of the β-D-xylosidase gene is shown in SEQ ID NO. 1, the transcript sequence of the β-D-xylosidase gene is shown in SEQ ID NO. 2, and the CDS sequence is shown in SEQ ID NO. 2, 201-2522 bp.

[0015] The beneficial effects are that the β-D-xylosidase gene of Caragana is cloned for the first time, and its biological functions of promoting root growth, sugar transport, cell wall morphology change, and stubble regeneration are clarified. Overexpression of the β-D-xylosidase gene has more vigorous growth ability, and the roots are longer than those of the wild type. After stubble, the β-D-xylosidase degrades polysaccharides in the root cell wall and transports upwards to provide energy for the regeneration process. The above functions of the β-D-xylosidase gene of Caragana are discovered for the first time, which provides a target gene for analyzing the stubble regeneration mechanism of Caragana and breeding high-quality Caragana. BRIEF DESCRIPTION OF DRAWINGS

[0016] Figure 1 The underlined part is the sequence of six exons in the full-length sequence of the β-D-xylosidase gene of Caragana.

[0017] Figure 2 The underlined part is the CDS sequence in the transcript sequence of the β-D-xylosidase gene of Caragana.

[0018] Figure 3 The underlined part is the CDS sequence in the transcript sequence of the β-D-xylosidase gene of Caragana.

[0019] Figure 4 The underlined part is the CDS sequence in the transcript sequence of the β-D-xylosidase gene of Caragana.

[0020] Figure 5 The underlined part is the CDS sequence in the transcript sequence of the β-D-xylosidase gene of Caragana.

[0021] Figure 6 The underlined part is the CDS sequence in the transcript sequence of the β-D-xylosidase gene of Caragana.

[0022] Figure 7 Figure 1 is a schematic diagram of Arabidopsis transformation. The figure includes the following steps: plant expression vector is transformed into Agrobacterium LBA4404, floral dip, resistance screening, plant transplanting culture, PCR detection of positive plants, etc.

[0023] Figure 8 Figure 2 is a phenotype analysis of β-D-xylosidase gene transgenic plants. The figure shows that the root length of the transgenic plants overexpressing β-D-xylosidase is significantly longer than that of wild type plants.

[0024] Figure 9 Figure 3 is a scanning electron microscope analysis of the root cell wall surface morphology of transgenic plants overexpressing β-D-xylosidase. Figure 9 a is a 20-micron scale wild type root cell wall surface morphology, Figure 9 b is a 10-micron scale wild type root cell wall surface morphology, Figure 9 c is a 20-micron scale transgenic plant overexpressing β-D-xylosidase root cell wall surface morphology, Figure 9 d is a 10-micron scale transgenic plant overexpressing β-D-xylosidase root cell wall surface morphology.

[0025] Figure 10 Figure 4 is a transmission electron microscope analysis of the root cell wall cross-sectional morphology of transgenic plants overexpressing β-D-xylosidase. Figure 10 a is a 2-micron scale wild type root cell wall surface morphology, Figure 9 b is a 1-micron scale wild type root cell wall surface morphology, Figure 9 c is a 2-micron scale transgenic plant overexpressing β-D-xylosidase root cell wall surface morphology, Figure 9 d is a 1-micron scale transgenic plant overexpressing β-D-xylosidase root cell wall surface morphology.

[0026] Figure 11 Figure 5 is a total sugar dynamic change analysis of transgenic plants overexpressing β-D-xylosidase. Figure 11 A is a sampling schematic diagram, CKSR is the root-stem junction of the control group, CKR is the middle part of the root of the control group, TSR is the root-stem junction of the stubble treatment group, and TR is the middle part of the root of the stubble treatment group, Figure 11 B is a comparison chart of total sugar content of each group. DETAILED DESCRIPTION

[0027] The methods and devices used in the following examples of the present application are all conventional methods and devices unless otherwise specified; the equipment and reagents used are all conventional equipment and reagents purchased from reagent companies. In order to make the purpose, technical scheme and advantages of the application clearer, the specific embodiments of the application will be described in detail below with reference to the drawings. The examples of these preferred embodiments are illustrated in the drawings. The embodiments of the application shown in the drawings and described according to the drawings are merely exemplary, and the application is not limited to these embodiments. Here, it should also be noted that, in order to avoid obscuring the technical scheme of the application due to unnecessary details, only the processing steps closely related to the scheme according to the application are shown in the drawings, and other details that are not closely related are omitted.

[0028] Example 1

[0029] The present embodiment provides a Caragana jubata β-D-xylosidase gene, the full-length sequence of which is shown in SEQ ID NO. 1 and Figure 1 as shown, including 7203bp, the underlined part in the figure is the exon sequence of the gene, and the unlined part is the intron and the upstream and downstream non-coding regions; the transcript sequence of the β-D-xylosidase gene is shown in SEQ ID NO. 2 and Figure 2 as shown, the underlined part in the figure is the sequence of the 2322bp CDS region.

[0030] Example 2

[0031] The present embodiment provides a method for screening Caragana jubata stubble regeneration key genes, which specifically comprises the following steps:

[0032] 1. Experimental material selection and transcriptome analysis

[0033] Caragana jubata planted in the Beishan experimental base in Yuzhong County, Lanzhou City, was selected for stubble experiment. Caragana jubata of the same habitat and age in the experimental base was selected for experiment, and the stubble experiment group was 100% stubble in mid-March. Leaf and root samples of the stubble experiment group and the control group were collected in early May, and 3 biological replicates of samples were collected for each group, and total RNA was extracted for transcriptome sequencing analysis. Transcriptome analysis was performed on the MGISEQ-T7 sequencing platform of Huada Gene, and general procedures were used for analysis, and non-parametric and parametric transcriptome analysis was performed at the same time. The specific method is referred to the website of Huada Gene and related literature (http: / / bgitechsolutions.com / sequencing / 45).

[0034] 2. Selection of stubble regeneration key genes

[0035] The genes with q value and p value less than 0.01 and log2FC greater than 4 were selected as the key genes in the process of pruning regeneration, and finally the key gene D-xylosidase gene was screened out, and the expression data is shown in Table 1.

[0036] Table 1 Expression of β-D-xylosidase gene screened by non-parametric transcriptome and parametric transcriptome analysis

[0037]

[0038] Example 3

[0039] The present embodiment provides an analysis method of Caragana β-D-xylosidase gene function, which specifically comprises the following steps:

[0040] 1. Cloning of Caragana β-D-xylosidase gene and construction of plant overexpression vector

[0041] (1) Selection of vector and medium

[0042] PRI201-AN plant overexpression vector, LB medium: 10 g / L peptone, 5 g / L yeast extract, 10 g / L NaCl, pH 7.0.

[0043] (2) Gene cloning

[0044] PCR reaction system: 2 μl 10x buffer, 2 μl 2 mM dNTPs, 2 μl DNA template, 1 μl DMSO, 0.8 μl 10 pmol / μl upper and lower primers (Table 2), 0.5 U kod DNA polymerase (KOD-401, TOYOBO), and water to 20 μl.

[0045] PCR amplification program: 94℃ pre-denaturation for 4 min; 94℃ denaturation for 30 s, 50-64℃ annealing for 30 s, 68℃ extension for 1 min / kb, 40 cycles; 68℃ incubation for 8 min.

[0046] Table 2 Primers for cloning of β-D-xylosidase gene

[0047]

[0048]

[0049] Electrophoresis: Add appropriate amount of 10x loading buffer to each reaction tube, and perform electrophoresis on 1%-3% (10ul EB, 1-3g agarose / 100ml 0.5x TBE buffer) agarose gel. Electrophoresis is carried out at 5-10 V / cm in 0.5x TBE buffer, and the electrophoresis is completed in a gel imaging system.

[0050] Agarose gel DNA recovery (using Jierui Biology GK2042 kit, according to the instructions): the DNA target band was carefully cut off and placed in a 1.5 ml EP tube. 400ul of banding B was added to the tube and placed in a 70℃ water bath until the gel was completely dissolved. 100ul of isopropyl alcohol was added to the tube, and it was placed at room temperature for 1 minute and centrifuged at 5000rpm for 1 minute to pass the column. Repeat the above steps once. Add 500ul of wash buffer 12000rmp wash twice, 10000rmp centrifuge 1 minute. Add 40ul of double distilled water to the column, place it at 37℃ for 2 minutes, centrifuge at 12000rmp for 1 minute to collect.

[0051] Double enzyme digestion: plasmid (30ul), 4ul 10x enzyme digestion buffer, 4ul 10x BSA, 6U restriction enzyme (NEB), add water to 40ul, treat in 37 degree water bath for 1h.

[0052] (3) Plasmid vector construction

[0053] As shown in Figure 6 , after the PCR fragment was recovered by agarose gel, it was mixed with the enzyme-digested recovered empty vector, and EasyGeno DNA recombination system was added (#VI201-02, Tiangen Biology). 10ul recombination system: 5ul 2x EasyGeno Assembly Mix, 2.5ul enzyme-digested vector DNA, 2.5ul fragment DNA. The reaction system was added to a 250ul EP tube, placed in a 50℃ water bath for 30 minutes, then transformed into E. coli and plated, placed in a 37℃ incubator for 16 hours, then picked bacteria and sent for sequencing. The correct plasmid was stored at -20℃ for long-term preservation.

[0054] Plasmid extraction: inoculate a single colony into 3 ml LB liquid medium containing appropriate antibiotics, 37°C, 200 rpm shaking culture overnight; take 1.5 ml of the culture (3 ml for low-copy plasmids), centrifuge at 12,000 rpm for 30 seconds; aspirate the supernatant, suspend the bacterial cells in 100 μl of solution I (Glucose 50 mmol / L, EDTA 10 mmol / L, Tris-HCl 25 mmol / L, pH 8.0); add 200 μl of freshly prepared solution II (NaOH 0.2 mol / L, SDS 1%), immediately mix gently; add 150 μl of solution III (KAc 5 mol / L, pH 4.8), mix quickly, and stand at room temperature for 5 minutes; centrifuge at 12,000 rpm for 10 minutes; transfer the supernatant to another centrifuge tube, add 2 volumes of ethanol, mix well; centrifuge at 12,000 rpm for 10 minutes; remove the supernatant, wash the DNA precipitate with 70% ethanol, centrifuge at 12,000 rpm for 1 minute, remove the supernatant; vacuum dry the precipitate; dissolve in 60 μl of double-distilled water containing 10 μg / ml RNase A.

[0055] 2. Analysis of the β-D-xylosidase gene structure of Caragana jubata

[0056] The β-D-xylosidase gene of Caragana jubata contains 7203 bp of nucleotides, including 6 exons ( Figure 1 and Figure 3 ); the transcript sequence is 2722 bp, including a CDS region of 2322 bp ( Figure 2 ). The β-D-xylosidase of Caragana jubata contains 773 amino acids, with amino acids 109-324 constituting the N-terminal Glyco-hydro-3 domain and amino acids 397-627 constituting the C-terminal Glyco-hydro-3 domain ( Figure 4 ). Homology BLAST search of the CDS sequence of the β-D-xylosidase gene of Caragana jubata was performed on NCBI, and the highest homologous sequence was downloaded to construct a homology phylogenetic tree, which showed that the β-D-xylosidase gene of Caragana jubata had low homology with genes in the NCBI database ( Figure 5 ).

[0057] 3. Transformation of the β-D-xylosidase gene of Caragana jubata into Arabidopsis thaliana Figure 7

[0058] (1) Planting of Arabidopsis thaliana

[0059] a. Preparation of the medium: the medium for Arabidopsis thaliana was selected as MS, which was prepared in advance (pH = 5.8).

[0060] ​b. Seed disinfection: add 1 ml of 5% sodium hypochlorite solution (containing 1 drop of Tween) to a 1.5 ml centrifuge tube, invert and mix for 8 minutes, rinse with sterile water for 5 times;

[0061] c. After disinfection, the seeds are divided and mixed with culture medium at 40-50°C, then poured into a flat plate, evenly covered with a layer (about 4-5 ml of culture medium is needed for a small culture dish);

[0062] d. Plate sealing, 2-3 days of vernalization in a 4°C refrigerator, then placed in an artificial climate chamber for germination and growth. The plant growth environment is 60% relative humidity, constant temperature 21-23°C, light cycle 16h light, 8h darkness, light intensity 80-200 μmol / M2 / S;

[0063] e. Preparation of planting soil: mix peat soil and vermiculite at a ratio of 2:1 and place for standby;

[0064] f. Soil immersion: fill the soil into the planting pot to about 1 cm from the pot opening, and completely immerse it with the compound fertilizer (N, P, K = 20%, 20%, 20%) of Hua Wuzhe;

[0065] g. Transplanting: 7-12 days after germination, select healthy and uniform growth seedlings and transplant them into the culture soil previously immersed with Hua Wuzhe, cover with plastic wrap, and remove the plastic wrap after the seedlings are alive.

[0066] (2) Agrobacterium culture

[0067] a. Take the Agrobacterium competent cells stored at -80°C to room temperature or hand for a while until they are partially thawed, and then insert them into ice when they are in an ice water mixed state.

[0068] b. Add 0.1 μg (volume not more than 10 μl) of plasmid DNA to each 100 μl of competent cells, mix well by tapping the tube bottom with hands, and then sequentially stand on ice for 5 minutes, liquid nitrogen for 5 minutes, 37°C water bath for 5 minutes, and ice bath for 5 minutes.

[0069] c. Add 700 μl of LB liquid medium without antibiotics, and cultivate at 28°C, 200 rpm for 2-3 hours.

[0070] d. Centrifuge at 6000 rpm for one minute to collect the bacteria, take about 100 μl of supernatant, gently blow and resuspend the bacterial block, and then spread on LB plate containing kanamycin 50 μg / ml, and invert and place in a 28°C incubator for 2-3 days. Randomly select one single colony, do colony PCR, and identify the correct Agrobacterium monoclonal for marking.

[0071] e. Use a sterile gun head to pick the marked Agrobacterium monoclonal and inoculate it into 1.5 ml of LB liquid medium containing the corresponding antibiotic (use a 50 ml blue cap centrifuge tube), and cultivate at 30 degrees, 200 rpm for 24 hours.

[0072] f. Inoculate the overgrown Agrobacterium culture into 100 ml LB liquid medium containing antibiotics at a ratio of 1%, and incubate at 30°C with shaking until OD600 = 1.0.

[0073] g. Centrifuge at 4,000 rpm for 15 min at 20°C to collect the bacterial cells.

[0074] h. Resuspend the bacterial cells in Transformation Buffer to OD600 = 1.0.

[0075] (3) Arabidopsis Transformation

[0076] a. Water the plants that are in the process of flowering one day in advance.

[0077] b. Invert the pots and immerse all the inflorescences in the bacterial suspension prepared in advance in Transformation Buffer for about 30 seconds.

[0078] c. Repeat the transformation as described above after 7 days. After 2-3 weeks, water the plants as little as possible to accelerate the aging process, and collect the mature seeds in a paper bag and dry them in a desiccator for 7 days.

[0079] (4) Screening of Transgenic Arabidopsis

[0080] a. Preparation of the medium: 1 / 2MS (0.8% agar powder, without sucrose, pH 5.8) is used for the medium of Arabidopsis.

[0081] b. Seed disinfection: 70% ethanol for 1 minute, 1 ml 7% sodium hypochlorite solution (containing 1 drop of Tween) for 10 minutes, invert and mix for 5 minutes, and rinse with sterile water for 5 times.

[0082] c. Resuspend the disinfected seeds with 100 μl sterile water, and use a 1 ml syringe to point them onto the 1 / 2MS medium (add selection antibiotics: 50 μg / ml KAN or 30 μg / ml HYG or 50 μM Glufosinate-ammonium) plates.

[0083] d. Seal the plates and vernalize them in a 4°C refrigerator for 48 hours, and then place them in an artificial climate chamber to start germination and growth. The plant growth environment is 60% relative humidity, constant temperature of 20-22°C, light cycle of 16 hours light and 8 hours darkness, and light intensity of 80-200 μmol / m2 / s. 2

[0084] e. Observe after 8-15 days, and transplant the positive plants into the planting soil.

[0085] f. Preparation of the planting soil: mix peat soil and vermiculite at a ratio of 2:1 and place for standby use.​

[0086] g. Soak the soil: Fill the planting pots with soil up to about 1 cm from the rim, and soak completely with Hanuoke compound fertilizer (N, P, K = 20%, 20%, 20%).

[0087] h. Transplanting: 20 days after germination, select healthy and uniform seedlings and transplant them into the soil soaked with Hanuoke, cover with plastic wrap, and remove after the seedlings are alive.

[0088] i. PCR identification: PCR detects the kanamycin resistance gene NPT, 670 bp, and the results show that 12 lines are positive. In the figure, Marker is 2000, 1000, 750 (brightest), 500, 250, 100 bp, and the sample order is marker, line 1-12, wild type control. Figure 7

[0089] 4. Phenotype and biochemical analysis of transgenic Arabidopsis

[0090] (1) β-D-xylosidase promotes root growth

[0091] As shown in Figure 8 , compared with the wild type, transgenic lines overexpressing β-D-xylosidase show better growth status and larger root length, indicating that the gene has a role in promoting plant growth and root development. β-D-xylosidase increases nutrient utilization, β-D-xylosidase promotes the decomposition of hemicellulose, which is the main component of plant cell wall. Improving carbon assimilation, xylose released by hemicellulose decomposition can be used as a carbon source for plants, and increased expression level of β-D-xylosidase may lead to faster release of xylose and subsequent plant assimilation, providing additional energy and carbon for the growth process. Enhance root configuration, the root length of transgenic lines indicates that the root configuration has changed, with the potential characteristics of increased branching or increased depth of roots into the soil. This improved root system can better absorb nutrients and water, promoting the overall growth and performance of the plant.

[0092] (2) β-D-xylosidase changes cell wall morphology

[0093] Overexpression of β-D-xylosidase gene in Arabidopsis has the potential for physiological and morphological changes. The main reason is the release of xylose by hemicellulose metabolism and cell wall remodeling. This leads to changes in cell wall, as well as changes in root growth and root hair. As shown in Figure 9 ​As shown in the root observation under scanning electron microscope, the cell wall porosity of the transgenic line roots was increased compared with the wild type. In the wild type, the root surface was smooth, while the root surface of the transgenic plants showed signs of degradation, with irregular pit formation, indicating that hemicellulose material was removed. This can change the permeability and ion exchange capacity of the root cell wall, potentially affecting the absorption and transport processes of nutrients. As shown in Figure 10 As shown in the transmission electron microscope observation, the wild type Arabidopsis root cell wall structure was complete, with large electron density and uniform thickness; while the root cell wall of the β-D-xylosidase gene overexpression Arabidopsis was uneven in thickness, with small electron density and varying degrees of degradation.

[0094] (3) β-D-xylosidase promotes sugar transport after mowing

[0095] As shown in Figure 11 As shown in A, the total sugar content of the control group and the mowing treatment for 3d of Arabidopsis root stem junction (Stem-root junction) and root middle (Root) tissue was determined. Due to the small sample size, 40 samples were mixed for each biological repeat, and a total of 120 samples were used in each group. The total sugar content was determined using the Shanghai Biosciences total sugar content detection kit (D799167) according to the instructions. As shown in Figure 11 As shown in B, the total sugar content of the root stem junction of the β-D-xylosidase gene overexpression line was significantly increased, and the total sugar content of the root middle was decreased, indicating that the total sugar in the root was transported to the root stem junction, and the enrichment of the total sugar content in the root stem junction was beneficial to the energy preparation for mowing regeneration.

[0096] The above is only a specific embodiment of the present application, and it should be noted that for ordinary skilled persons in the art, without departing from the principles of the present application, a number of improvements and refinements can be made, which should also be considered as the protection scope of the present application.

Claims

1. The application of overexpression of the β-D-xylosidase gene to promote Arabidopsis root growth, characterized in that... The full-length sequence of the β-D-xylosidase gene is shown in SEQ ID NO.1, and the transcript sequence of the β-D-xylosidase gene is shown in SEQ ID NO.2.

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