Application of cytokinin response factor RhARR17 in regulating axillary bud germination of Rosa chinensis

Abstract

The present invention relates to the field of biotechnology, and particularly to a cytokinin response factor RhARR17 and its application in regulating the axillary bud germination of roses. The present invention provides the application of a product that inhibits RhARR17 the expression of a gene or its transcription and translation proteins in promoting the axillary bud germination of roses. The base sequence of the RhARR17 gene is as shown in SEQ ID NO.1. In the present invention, according to the RhARR17 expression results in buds at different sites of rose flower branches, RhARR17 was transiently silenced in axillary buds, and it was found that RhARR17 abnormal expression would affect the germination and growth of axillary buds, and silencing RhARR17 could promote the growth of axillary buds; on the contrary, transient overexpression of RhARR17 in axillary buds could inhibit the growth of axillary buds.

Description

Technical Field

[0001] The present invention relates to the field of biotechnology, and particularly to the application of the RhARR17 gene in regulating the germination of axillary buds of roses. Background Art

[0002] Rose (Rosa hybrida) is a perennial evergreen or semi-evergreen woody ornamental plant of the genus Rosa in the family Rosaceae. Roses have the characteristics of elegant flower shapes, bright colors, and rich fragrances, and have high ornamental and economic values. They rank first among the world's four major cut flowers and are one of the most consumed flowers in the cut flower market. They are widely used in landscaping, courtyard decoration, potted flowers, perfume, and cosmetics industries.

[0003] Germination is an important biological characteristic in the process of plant growth and development. The germination of axillary buds determines the formation of plant branches, and the number of branches directly affects the number of vegetative organs, thus having a decisive impact on crop yields. The differentiation process of meristems forms axillary buds. First, the meristems differentiate into lateral bud primordia, which further develop into axillary buds and finally form lateral branches. The number of plant lateral branches is closely related to the germination of axillary buds, and the two show a positive correlation.

[0004] The axillary buds of roses mainly develop in the leaf axils, and their further growth can form flowering branches. Therefore, this process is crucial for the growth and morphogenesis of roses, and axillary buds are also one of the most important propagation materials for roses. In the production process of cut roses, the germination rate of axillary buds directly affects their survival rate; during the entire growth and development process, the characteristics of axillary buds determine the growth cycle, yield, and quality of cut roses. Therefore, studying and regulating the related factors of rose axillary bud germination has important practical significance for the breeding of cut roses and the improvement of industrial quality and efficiency. Summary of the Invention

[0005] The present invention screened out the cytokinin response factor RhARR17 and found that this gene is highly expressed in the axillary buds and sepals of rose tissues, affects the germination of rose axillary buds, and inhibiting the expression of this gene can promote the germination of rose axillary buds.

[0006] To achieve the above objectives, the present invention provides the following technical solutions:

[0007] On the one hand, the present invention provides the application of inhibiting the expression of the RhARR17 gene or its transcription and translation proteins in promoting the germination of rose axillary buds, and the base sequence of the RhARR17 gene is as shown in SEQ ID NO.1.

[0008] On the other hand, the present invention provides an application of a product that inhibits the expression of the RhARR17 gene or its transcription and translation protein in promoting the germination of axillary buds of roses, and the base sequence of the RhARR17 gene is as shown in SEQ ID NO.1.

[0009] Furthermore, the product is a product that silences, knocks out or knocks down the RhARR17 gene.

[0010] Furthermore, the product that silences the RhARR17 gene contains the base sequence shown in SEQ ID NO.2.

[0011] On the other hand, the present invention provides an application of a kit in promoting the germination of axillary buds of roses, the kit contains a reagent that inhibits the expression of the RhARR17 gene, and the base sequence of the RhARR17 gene is as shown in SEQ ID NO.1.

[0012] On the other hand, the present invention provides an isolated polynucleotide, the base sequence of the polynucleotide is as shown in SEQ ID NO.2, and the polynucleotide is used to inhibit the expression of the RhARR17 gene or its transcription and translation protein, and the base sequence of the RhARR17 gene is as shown in SEQ ID NO.1.

[0013] On the other hand, the present invention provides a vector, and the vector contains the base sequence shown in SEQ ID NO: 2.

[0014] On the other hand, the present invention provides an engineered bacterium, and the engineered bacterium contains the above-mentioned vector.

[0015] Furthermore, the present invention provides an application of the above-mentioned polynucleotide, or the above-mentioned vector, or the above-mentioned engineered bacterium in promoting the germination of axillary buds of roses.

[0016] On the other hand, the present invention provides a method for promoting the germination of axillary buds of roses, the method is to inhibit the expression of the RhARR17 gene or its transcription and translation protein in roses, and the base sequence of the RhARR17 gene is as shown in SEQ ID NO.1.

[0017] Advantages of the present invention:

[0018] The present invention identified a cytokinin response factor RhARR17 from the rose genome, and found that this gene is highly expressed in axillary buds and sepals of rose tissues, the expression signal is concentrated in axillary buds, and there is almost no expression in petals and stem segments. In the analysis of the expression of RhARR17 in buds at different positions of rose flower branches, it was found that the expression of RhARR17 is high in axillary buds in the middle part. Therefore, the present application speculates that RhARR17 may be involved in the germination of axillary buds of roses.

[0019] In this invention, according to the expression results of RhARR17 in buds at different positions of rose flower branches, RhARR17 in axillary buds was transiently silenced, and it was found that the abnormal expression of RhARR17 would affect the germination and growth of axillary buds. Silencing RhARR17 could promote the growth of axillary buds; transient overexpression of RhARR17 in axillary buds could inhibit the growth of axillary buds. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 is the expression analysis diagram of RhARR17 provided by the present invention; Note: They are stem segments, roots, petals, leaves, sepals, axillary buds, stamens, and pistils respectively.

[0021] Figure 2 is the subcellular localization diagram of RhARR17 provided by the present invention; Note: (A) Prediction result of subcellular localization of RhARR17; (B) Subcellular localization in Nicotiana benthamiana leaves. RhARR17-GFP and NF-YA4-mCherry (nuclear marker) were co-infiltrated into tobacco leaves, and fluorescence signals were observed by confocal microscopy.

[0022] Figure 3 is the statistical chart of quantitative analysis of bud points at different nodes of RhARR17; Data are shown as mean ± SD (n = 3) (**** indicates P ≤ 0.0001), which are upper axillary buds, middle axillary buds, and lower axillary buds respectively.

[0023] Figure 4 is the phenotypic observation result diagram of axillary bud germination and growth at different time periods after transient silencing of RhARR17 provided by the present invention; Note: 0d, 4d, 8d represent the growth conditions of axillary buds silenced by TRV2 and TRV2-RhARR17 on the 0th, 4th, and 8th days after infection.

[0024] Figure 5 is the anatomical diagram of axillary bud growth status at different time periods after transient silencing of RhARR17 provided by the present invention; Note: 0d, 4d, 8d represent the stereoscopic anatomical observation conditions of the growth points of axillary buds silenced by TRV2 and TRV2-RhARR17 on the 0th, 4th, and 8th days after infection.

[0025] Figure 6 is the statistical chart of gene expression level and axillary bud length measurement after transient silencing of RhARR17 provided by the present invention; Note: (A) RT-qPCR analysis of the relative expression level of RhARR17 after silencing; (B) Bud lengths of the control group TRV2 and the treatment group TRV2-RhARR17 silenced at 0d, 4d, and 8d after silencing (* indicates P ≤ 0.05, ** indicates P ≤ 0.01).

[0026] Figure 7It is the phenotypic observation result diagram of axillary bud germination and growth at different time periods after transient overexpression of RhARR17 provided by the present invention; Note: 0d, 6d, and 12d represent the growth conditions of axillary buds overexpressed by pSuper-1300 and pSuper-1300-RhARR17 on the 0th, 6th, and 12th days after infection.

[0027] Figure 8 It is the anatomical diagram of the growth state of axillary buds at different time periods after transient overexpression of RhARR17 provided by the present invention; Note: 0d, 6d, and 12d represent the stereoscopic anatomical observation conditions of the growth points of axillary buds overexpressed by Super-1300 and pSuper-1300-RhARR17 on the 0th, 6th, and 12th days after infection.

[0028] Figure 9 It is the statistical chart of gene expression level and axillary bud length measurement after transient overexpression of RhARR17 provided by the present invention; Note: (A) RT-qPCR analysis of the relative expression level of RhARR17 after overexpression; (B) Bud lengths of the control group pSuper-1300 and the treatment group pSuper-1300-RhARR17 after overexpression at 0d, 6d, and 12d (* indicates P≤0.05, ** indicates P≤0.01). Detailed implementation manners

[0029] The following is a detailed description in combination with examples and drawings. In this application, transcriptome sequencing was performed on axillary buds at different time nodes after pruning, and a cytokinin response factor RhARR17 with significant differences related to rose axillary bud germination was screened out. It is highly expressed in axillary buds, and it is found that the expression level of RhARR17 is related to axillary bud activity, with the highest expression in upper axillary buds, followed by middle axillary buds, and the lowest expression in lower axillary buds. After transient silencing and transient overexpression of RhARR17, it was found that the germination and growth rate of axillary buds were affected. Therefore, this application reveals the biological function of RhARR17 in rose axillary bud germination, which is of extremely important significance for cut rose breeding and industrial quality improvement and efficiency increase.

[0030] The base sequence of the RhARR17 gene provided by the present invention is shown in SEQ ID NO: 1, and the base sequence of the silenced RhARR17 gene is shown in SEQ ID NO: 2.

[0031] The abbreviations and their corresponding names that appear in this application are shown in Table 1.

[0032] Table 1 Abbreviation table

[0033] Example

[0034] 1. Plant materials

[0035] 1.1 ‘Pink Snow Mountain’

[0036] ‘Pink Snow Mountain’ is a modern cut rose variety, which is taken from the Baofeng Base of the Institute of Floriculture, Yunnan Academy of Agricultural Sciences. The single-bud stem segments are used as materials for the axillary bud germination experiment of roses.

[0037] 1.2 Tobacco

[0038] The experimental material for subcellular localization is Nicotiana benthamiana. The seeds are sown in a moist nutrient substrate, covered with a film, and cultured in a culture room.

[0039] Cultivation conditions: temperature 24 ± 1 °C, relative humidity 60 - 65%, photoperiod 16 h / 8 h.

[0040] 1.3 Strains and vectors

[0041] Escherichia coli: DH5α and Agrobacterium tumefaciens strains: EHA105, pSuper-1300 (Kan resistance) are all purchased from Beijing Tsingke Biotechnology Co., Ltd. The VIGS vectors are pTRV1 and pTRV2 purchased from HonorGene.

[0042] 1.4 Medium formulations involved in the case

[0043] (1) LB medium and YEB medium (Table 2)

[0044] Table 2 Formulations of LB and YEB media

[0045]

[0046] 2. Research methods

[0047] 2.1 Extraction of total RNA

[0048] Use the RNAprep Pure Polysaccharide and Polyphenol Plant Total RNA Extraction Kit (centrifugal column type) to extract total RNA from the petals, leaves, receptacles, sepals, and axillary buds of ‘Pink Snow Mountain’.

[0049] 2.2 cDNA synthesis

[0050] (1) Removal of genomic DNA (Table 3)

[0051] Table 3 Reaction system for removal of genomic DNA

[0052]

[0053] Mix well, 42 °C, 2 min.

[0054] (2) Prepare the reverse transcription reaction system (Table 4)

[0055] Table 4 Transcription reaction system

[0056]

[0057] Pipette and mix well, incubate at 37 °C for 15 min; incubate at 85 °C for 5 s. Store the product at -20 °C.

[0058] 2.3 Real-time fluorescence quantitative PCR

[0059] Design specific primers for the gene using Primer premier5 (Table 5). Dilute the cDNA four-fold with ddH 2 O, and use the reverse-transcribed cDNA as a template for Real-Time PCR amplification. Use GAPDH as an internal reference, and set up 3 biological replicates.

[0060] Table 5 RT-qPCR primer sequences

[0061]

[0062] The reaction system is shown in Table 6:

[0063] Table 6 RT-qPCR reaction system

[0064]

[0065] The reaction program is shown in Table 7:

[0066] Table 7 RT-qPCR reaction program

[0067]

[0068] 2.4 Vector construction

[0069] 2.4.1 PCR amplification of the target gene fragment

[0070] Use a high-fidelity enzyme (PhusionTM Plus PCR Master Mix) for PCR amplification of the target gene. The reaction system is shown in Table 8:

[0071] Table 8 PCR amplification reaction system

[0072]

[0073] The reaction program is shown in Table 9:

[0074] Table 9 PCR amplification reaction program

[0075]

[0076] After the amplification, perform 1% gel electrophoresis and select the target band for subsequent gel extraction.

[0077] 2.4.2 Gel Extraction

[0078] Use a DNA gel extraction kit (TaKaRa MiniBEST Agarose Gel DNA Extraction Kit Ver.4.0) to recover and purify the PCR products.

[0079] 2.4.3 Double Digestion of the Vector

[0080] According to the restriction enzyme sites for the insertion of the target fragment sequence, perform double digestion of the vector with the corresponding enzymes. The double digestion system is shown in Table 10:

[0081] Table 10 Double Digestion System

[0082]

[0083] After adding the reagents of the system on ice, perform double digestion of the vector according to the heat denaturation temperature of the specific enzyme.

[0084] 2.4.4 Homologous Recombination

[0085] Perform homologous recombination of the double-digested vector and the cloned target gene fragment to construct the vector. The homologous recombination system is shown in Table 11:

[0086] Table 11 Homologous Recombination System

[0087]

[0088] Procedure: Run at 50 °C for 15 min in a PCR instrument.

[0089] 2.4.5 Transformation of Escherichia coli

[0090] (1) Take out the competent cells from -80 °C and thaw them on ice;

[0091] (2) Take a 1.5 mL centrifuge tube, add 10 μL of the recombinant product and 50 μL of DHα competent cells, pipette and mix well, and let it stand on ice for 30 min;

[0092] (3) Heat shock in a 42 °C water bath for 90 s, and quickly transfer it to ice and let it stand for 2 min;

[0093] (4) Add 500 μL of LB to the centrifuge tube, incubate at 37 °C, 200 rpm for 1 h;

[0094] (5) Centrifuge the cultured bacterial solution at 5000 rpm for 5 min;

[0095] (6) Discard 400 μL of the supernatant in a laminar flow hood. After pipetting and mixing evenly, spread it on an LB solid medium containing antibiotics and incubate it overnight at 37 °C in an inverted position.

[0096] 2.4.6 Bacterial Inspection and Sequencing

[0097] (1) Shake the bacteria: Pick a single colony grown overnight and culture it in 500 μL of LB medium containing antibiotics by shaking for 3 - 4 h (37 °C, 200 rpm).

[0098] (2) Bacterial liquid PCR: The cultured bacterial liquid is used for PCR amplification to detect whether the target band of the constructed vector meets the expectation. The bacterial liquid PCR amplification system is shown in Table 12:

[0099] Table 12 Bacterial Liquid PCR Amplification System

[0100]

[0101] The bacterial liquid PCR amplification program is shown in Table 13:

[0102] Table 13 Bacterial Liquid PCR Amplification Reaction Program

[0103]

[0104] The amplification product is used for 1% gel electrophoresis imaging, and the samples with the band size meeting the expectation are sent to the company for sequencing.

[0105] 2.5 Plasmid Extraction

[0106] After the sequencing results are returned, perform sequence alignment, select the samples that meet the expectation, shake the bacteria, and extract the plasmid using a plasmid extraction kit (TaKaRa MiniBEST Plasmid Purification Kit Ver.4.0).

[0107] 2.6 Agrobacterium Transformation

[0108] (1) Take out the Agrobacterium competent cells from the -80 °C refrigerator. When they are melted to the state of an ice - water mixture, insert them into ice.

[0109] (2) Add 0.01 - 1 μg of plasmid DNA to every 100 μL of competent cells. After pipetting and mixing evenly, let it stand on ice for 5 min, in liquid nitrogen for 5 min, in a 37 °C water bath for 5 min, and in an ice bath for 5 min in sequence.

[0110] (3) Add 500 μL of antibiotic - free YEB liquid medium and culture it by shaking at 28 °C for 2 - 3 h.

[0111] (4) Centrifuge at 5000 rpm for 2 min. In a laminar flow hood, discard 400 μL of the supernatant, and spread it onto YEB solid medium containing antibiotics. Incubate the plate upside down at 28 °C for 2 - 3 d.

[0112] 2.7 Subcellular localization

[0113] (1) Vector construction

[0114] Select SmaI and KpnI as restriction sites, insert the complete coding region (SEQ ID NO: 1) sequence of RhARR17 into the pSuper - 1300 vector, design primers using the homologous recombination method to construct the RhARR17 - GFP vector. After obtaining the Agrobacterium positive strain, perform the following experiments.

[0115] (2) Bacterial shaking culture

[0116] 1) Streak culture of the bacterial solution: Take out the stored bacterial solution and streak it on YEB medium containing the corresponding antibiotics. Incubate the plate upside down at 28 °C for 2 - 3 days;

[0117] 2) Bacterial shaking culture: Pick a single colony into a 1.5 mL centrifuge tube, add 500 μL of YEB liquid medium containing antibiotics for overnight small - scale shaking (28 °C, 200 rpm), perform PCR bacterial detection. For the bacterial solution with correct bands, continue with medium - scale and large - scale shaking.

[0118] (3) Tobacco injection

[0119] 1) Bacterial collection: Centrifuge the bacterial solution cultured to OD 600 = 0.4 - 0.6 (5000 rpm, 8 min), discard the supernatant, and collect the bacterial cells. Resuspend the bacterial cells with the infection solution, adjust the OD 600 to 1.0, and mix according to the ratio of RhARR17 - GFP / GFP:NF - YA4 - mCherry:P19 = 1:1:0.5. Let it stand in the dark for 3 h.

[0120] 2) Injection: After standing, gently make a cut on the back of the tobacco (grown to 4 - 6 leaves) with a needle, and use a 1 mL syringe to inject the proportionally mixed bacterial solution into the tobacco leaves, and make marks;

[0121] 3) Incubation: Place the injected tobacco in an incubator at 25 °C, first incubate it in the dark for one day, and then transfer it to light for 2 days; During this period, pay attention to watering to keep the tobacco growing normally.

[0122] (4) Observation and photography

[0123] Select the leaves near the tobacco injection port, gently cut them into squares with a blade and forceps, place them on a glass slide, drop distilled water on them, and cover them with a coverslip (pay attention to avoiding the generation of air bubbles). Observe and take pictures with the laser confocal microscope of the State Key Laboratory of the Institute of Biology, Yunnan Academy of Agricultural Sciences.

[0124] 2.8 Transient silencing

[0125] (1)Vector primer design and vector construction

[0126] Using EcoRI and BamHI as restriction enzyme sites, insert the silencing fragment (SEQ ID NO: 2) sequence of RhARR17 into the empty TRV2 vector. Design primers by the method of homologous recombination to construct the TRV-RhARR17 vector.

[0127] (2)Bacterial liquid culture

[0128] Streak the bacterial liquid on a plate (containing 50 mg / L Kan / Rif), and incubate it upside down at 28 °C for 2 - 3 days. Pick a single colony and shake it gently in 500 μL of YEB containing antibiotics for bacterial inspection; for those with correct bands, perform medium shaking and large shaking (28 °C, 200 rpm).

[0129] (3)Bacterial collection and resuspension

[0130] Centrifuge to collect bacteria at 5000 rpm for 8 min, pour off the supernatant, resuspend the bacterial cells with the infection solution, pipette and mix well, and adjust to OD600 = 1.0. When performing the transient silencing experiment, mix the TRV1 and TRV2, and TRV2-RhARR17 bacterial solutions in a volume ratio of 1:1, and let it stand in the dark for 4 - 6 h.

[0131] (4)Vacuum aspiration

[0132] Use a vacuum pump to aspirate and infect the stem segments with single buds at 0.082 MPa for 10 min, hold the pressure for 10 min, and release the air for 10 min, and try to immerse the entire stem segment in the bacterial solution as much as possible, and repeat the treatment three times. After infection, rinse with sterile water, place it in an 8 °C culture room for 3 d and then cuttage, observe every two days, and take samples and take pictures for record.

[0133] 2.10 Primers used in the RhARR17 sequence experiment

[0134] Table 14 RhARR17 sequence and primer list

[0135]

[0136] 3 Experimental results

[0137] 3.1 Expression verification of RhARR17 in different tissue parts

[0138] Total RNA was extracted from the petals, leaves, receptacles, sepals, and axillary buds of roses in this application, and its quality was detected. Genes with the most cis - acting elements related to meristems and plant growth and development were selected. Fluorescent quantitative expression analysis found that the expression of different genes varied in different tissues ( Figure 1 ). Among them, RhARR17 was specifically and highly expressed in axillary buds and sepals, indicating that RhARR17 might be crucial for the growth and development of rose axillary buds.

[0139] 3.2 Subcellular localization analysis of RhARR17

[0140] To explore the subcellular location where RhARR17 functions, this application first performed an online prediction using Cell - PLoc (http: / / www.csbio.sjtu.edu.cn / bioinf / Cell - PLoc - 2), which showed localization in the nucleus ( Figure 2 A). To further verify the accuracy of the localization, the CDS sequence of RhARR17 was cloned to construct the RhARR17 - GFP vector. Using the GFP empty vector as a control, after transferring it into Agrobacterium EH105, the leaves of Nicotiana benthamiana were injected. After 3 days, observation was carried out under a laser confocal microscope, and the results showed that both the GFP empty vector and RhARR17 - GFP were localized in the nucleus ( Figure 2 B).

[0141] 3.4 Analysis of the expression level of RhARR17 in different active buds

[0142] This application performed real - time fluorescence quantitative PCR analysis on the expression level of RhARR17 in axillary buds at different node positions of the cut rose 'Pink Avalanche' (the flower buds were not showing white). It was found that the expression of RhARR17 was relatively high in the upper axillary buds, followed by the middle axillary buds, and the lowest in the lower axillary buds ( Figure 3 ). Therefore, this application speculated that RhARR17 was closely related to the germination of axillary buds.

[0143] 3.5 RhARR17 inhibits the germination of rose axillary buds

[0144] Based on the analysis results of the expression level of RhARR17 in buds at different sites, this application selected a specific segment of the RhARR17 sequence (SEQ ID NO: 2) and used homologous recombination to construct a TRV - RhARR17 transient silencing vector. After transferring TRV1, TRV2, and TRV2 - RhARR17 into Agrobacterium EHA105, it was used to infect the stem segments of middle axillary buds to silence RhARR17, and phenotypic observations and records were carried out at 0 d, 4 d, and 8 d after infection ( Figure 4). From the phenotypic perspective of the buds, compared with the TRV2 control, the germination rate of the TRV2-RhARR17 buds was faster. The anatomical results of the buds also showed that the development of the buds accelerated after RhARR17 was silenced ( Figure 5 ). To confirm the silencing effect, total RNA of the buds before and after RhARR17 silencing was extracted, and RT-qPCR analysis was performed on its expression level. It was found that the expression decreased extremely significantly after RhARR17 silencing, indicating that the silencing was effective ( Figure 6 A). In addition, the bud lengths of the control group TRV2 and the experimental group TRV2-RhARR17 at different time points were also measured, and it was found that the difference was extremely significant on the 10th day after silencing ( Figure 6 B).

[0145] Based on the analysis results of the expression level of RhARR17 in buds at different sites, the CDS sequence (SEQ ID NO:1) of the RhARR17 sequence was selected in this application. A transient overexpression vector pSuper-1300-RhARR17 was constructed by homologous recombination method. After transferring pSuper-1300 and pSuper-1300-RhARR17 into Agrobacterium EHA105, they were used to infect the stem segments of the middle axillary buds to overexpress RhARR17. Phenotypic observations and records were carried out at 0 d, 6 d, and 12 d after infection ( Figure 7 ). From the phenotypic perspective of the buds, compared with the pSuper-1300 control, the germination rate of the pSuper-1300-RhARR17 buds was slower. The anatomical results of the buds also showed that the development of the buds slowed down after RhARR17 was overexpressed ( Figure 8 ). To confirm the overexpression effect, total RNA of the buds before and after RhARR17 overexpression was extracted, and RT-qPCR analysis was performed on its expression level. It was found that the expression increased extremely significantly after RhARR17 overexpression, indicating that the overexpression was effective ( Figure 9 A). In addition, the bud lengths of the control group pSuper-1300 and the experimental group pSuper-1300-RhARR17 at different time points were also measured, and it was found that the difference was extremely significant on the 12th day after silencing ( Figure 9 B).

[0146] It should be noted that the above specific embodiments are exemplary. Those skilled in the art can come up with various solutions inspired by the disclosed content of the present invention, and these solutions also fall within the scope of the disclosure of the present invention and within the protection scope of the present invention. Those skilled in the art should understand that the description and drawings of the present invention are illustrative and do not constitute a limitation on the claims. The protection scope of the present invention is defined by the claims and their equivalents.

Claims

1. Application of inhibiting the expression of RhARR17 gene or its transcriptional translation protein in promoting the germination of axillary buds of rose, characterized in that: The base sequence of the RhARR17 gene is shown in SEQ ID NO.

1.

2. The use of a product for inhibiting the expression of the RhARR17 gene or its transcriptional translation protein in promoting the germination of axillary buds of roses, characterized in that: The base sequence of the RhARR17 gene is shown in SEQ ID NO.

1.

3. The use according to claim 2, characterized in that: The product is a product for silencing, knocking out or knocking down the RhARR17 gene.

4. The use according to claim 3, characterized in that: The product for silencing the RhARR17 gene contains the base sequence described in SEQ ID NO.

2.

5. Use of a kit in promoting the germination of axillary buds of roses, characterized in that: The kit contains a reagent for inhibiting the expression of the RhARR17 gene, and the base sequence of the RhARR17 gene is shown in SEQ ID NO.

1.

6. An isolated polynucleotide, characterized in that The base sequence of the polynucleotide is shown as SEQ ID NO.

2. The polynucleotide is used to inhibit the expression of the RhARR17 gene or its transcriptional translation protein. The base sequence of the RhARR17 gene is shown as SEQ ID NO.

1.

7. A carrier, characterized in that The vector contains the base sequence shown in SEQ ID NO:

2.

8. An engineered bacterium, characterized in that: The engineered bacteria contains the vector according to claim 7.

9. Use of the polynucleotide according to claim 6, or the vector according to claim 7, or the engineered bacteria according to claim 8 in promoting the germination of axillary buds of roses.

10. A method for promoting the germination of axillary buds of roses, characterized in that: The expression of the RhARR17 gene or its transcriptional translation protein in rose is inhibited, and the base sequence of the RhARR17 gene is shown in SEQ ID NO.1.

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