Grape VvKCS11 promoter and application thereof

Abstract

The invention provides a grape VvKCS11 promoter and application thereof, belongs to the technical field of plant genetic engineering, and particularly provides a promoter ProKCS11 responding to salt stress, and the nucleotide sequence of the promoter ProKCS11 is shown as SEQ ID NO.1; a recombinant vector, an expression cassette, a recombinant cell or a recombinant bacterium containing the ProKCS11 promoter with the nucleotide sequence as shown in SEQ ID NO. 1; the promoter ProKCS11 has the effect of responding to salt stress, drought stress, osmotic stress, ion stress or abscisic acid, salt stress, drought stress and ion toxicity can induce the promoter to drive high expression of downstream genes, and the expression has certain tissue specificity; vvKCS11 overexpression driven by ProKCS11 can improve the stress resistance of the transgenic plant.

Description

Technical Field

[0001] The present invention belongs to the technical field of plant genetic engineering, and particularly relates to a grape VvKCS11 promoter and its application. Background Art

[0002] Soil salinization is one of the major problems threatening agricultural production and ecological environment, seriously threatening crop yields and the sustainable development of agriculture. Grape ( Vitis vinifera L.) is one of the important economic fruit trees, with high economic value and nutritional value. The distribution of saline soil in China involves many grape-producing areas. Although grapes have certain salt tolerance, their growth is limited and their yields are reduced under high-salt conditions, seriously restricting the sustainable development of the grape industry in China.

[0003] Under salt stress, the cork layer of grape roots thickens and its occurrence position is closer to the root tip, which can effectively prevent Na + and Cl - from being transported to the stele through the apoplastic flow. β-Ketoacyl CoA Synthase (KCS) participates in the fatty acid chain elongation reaction and is the rate-limiting enzyme for the synthesis of suberin precursors. Research shows that grape VvKCS11 is related to the formation of the root cork layer, and overexpression of this gene can significantly improve the salt tolerance of transgenic Arabidopsis thaliana.

[0004] CaMV35S Constitutive promoters such as VvKCS11 have the characteristics of simple operation and stable expression, and have played an important role in the research of plant stress-resistant gene functions for many years. However, although the stable expression of stress-resistant genes in the whole plant can improve the stress resistance of plants, it often leads to varying degrees of growth retardation, which limits the application of stress-resistant genes in breeding. The differences in promoters of plant stress-resistant related genes are an important reason for the differences in resistance among varieties during the process of plant evolution and domestication. These promoters are induced by salt stress, and the downstream genes driven by them often affect the salt tolerance of plants through tissue-specific expression. Existing research has determined the VvKCS11 role in plant salt tolerance, but the verification of this function was carried out through the constitutive promoter CaMV35S and the research on the VvKCS11 promoter is still blank. Whether this promoter will affect the VvKCS11 salt resistance effect in plants is not clear. Summary of the Invention

[0005] Aiming at the deficiencies of the prior art, the present invention provides a grape VvKCS11 promoter and its application.

[0006] The technical solution of the present invention is as follows: A promoter responsive to salt stress, drought stress, osmotic stress, ionic stress or abscisic acid ProKCS11 , said promoter ProKCS11 has the nucleotide sequence shown in SEQ ID NO.1.

[0007] A biological material, said biological material being a recombinant vector, expression cassette, recombinant cell or recombinant bacterium containing the promoter with the nucleotide sequence shown in SEQ ID NO.1 ProKCS11 .

[0008] The above promoter ProKCS11 or the above biological material is used in any of the following: ① Preparing a transgenic plant breeding preparation for stress resistance; ② Improving the stress resistance of plants; ③ Cultivating transgenic plants for stress resistance.

[0009] Preferably according to the present invention, the stress resistance in the above application is salt stress resistance, drought stress resistance, osmotic stress resistance or ionic stress resistance.

[0010] Preferably according to the present invention, the above application is to express a target gene in a plant through a promoter ProKCS11 .

[0011] More preferably, the target gene is a stress resistance gene.

[0012] More preferably, the stress resistance genes include salt tolerance genes and drought resistance genes.

[0013] More preferably, the stress resistance gene is VvKCS11 gene.

[0014] More preferably, the VvKCS11 gene has the nucleotide sequence shown in SEQ ID NO.2.

[0015] Preferably according to the present invention, the plant is grape.

[0016] Beneficial effects 1. The promoter provided by the present invention ProKCS11 has the function of responding to salt stress, drought stress, osmotic stress, ionic stress or abscisic acid. Under salt stress conditions, the promoter ProKCS11 can enhance the expression of the gene it drives, and this induction can occur under osmotic stress and ionic toxicity equivalent to the salt stress, and the gene expression it drives has a certain tissue specificity.

[0017] 2. In the present invention, the promoter ProKCS11 drives VvKCS11Overexpressed transgenic grape calli showed better salt tolerance, with growth rate, membrane stability, ion homeostasis, etc. being superior to those of the wild type in many aspects. Description of the Drawings

[0018] Figure 1 For VvKCS11 PCR amplification diagram of the gene promoter in grapes; In the figure: Lanes 1-3: VvKCS11 Promoter ProKCS11 Amplified fragment; M: DL2000 DNA Marker.

[0019] Figure 2 For the construction of the plant expression vector ProKCS11 :: GUS-1300GN and the verification diagram of Agrobacterium colony PCR; In the figure: (a) Double digestion of pBlunt- ProKCS11 and 1300GN vectors with SalI and BamHI; (b) ProKCS11 ::GUS-1300GN Agrobacterium colony PCR; M: DL2000 DNA Marker; Lanes 1-2: pBlunt- ProKCS11 Double digestion fragment; Lane 3: 1300GN vector SalI and BamHI double digestion fragment; Lanes 4-10: ProKCS11 :: GUS-1300GN Agrobacterium colony PCR positive fragment.

[0020] Figure 3 For the screening diagram of transgenic Arabidopsis thaliana on 1 / 2 MS + hygromycin medium.

[0021] Figure 4 For the PCR identification diagram of transgenic Arabidopsis thaliana; In the figure: M: DL2000 DNA Marker; Lanes 1-4: PCR products of 4 resistant screening positive Arabidopsis thaliana plants.

[0022] Figure 5 For the GUS staining analysis diagram of transgenic Arabidopsis thaliana seedlings after 48 hours of treatment under different stress conditions; In the figure: The control is the ProKCS11 :: GUS-1300GN transgenic Arabidopsis thaliana plants without any stress treatment.

[0023] Figure 6 For the construction of the plant expression vector ProKCS11 :: VvKCS11- pCLB1301 and the verification diagram of Agrobacterium colony PCR; In the figure: (a) Double digestion diagram of the vector and the target fragment, Lanes 1-2: ProKCS11-pEASY Blunt Vector digestion diagram; Lanes 3-4:VvKCS11-pCLB Vector digestion map; (b) ProKCS11 :: VvKCS11- Agrobacterium tumefaciens colony PCR of pCLB1301. Lanes 1-8 show the PCR results of 8 single colonies; M1: DL5000 DNA Marker; M2: DL2000 DNA Marker.

[0024] Figure 7 Verification map of transgenic grape callus; In the figure: A. Wild-type grape callus under bright field; B. Under bright field ProKCS11 :: VvKCS11- pCLB1301 overexpressed grape callus; C. Wild-type grape callus under fluorescence excitation; D. Under fluorescence excitation ProKCS11 :: VvKCS11- pCLB1301 overexpressed grape callus.

[0025] Figure 8 Phenotype map of wild-type and ProKCS11 :: VvKCS11- pCLB1301 overexpressed grape callus after 14 days of treatment with 0 mM and 100 mM NaCl;

[0026] Figure 9 Ion change map of wild-type and ProKCS11 :: VvKCS11- pCLB1301 overexpressed grape callus after 14 days of treatment with 0 mM and 100 mM NaCl; In the figure: A. Na + content change map; B. K + content change map; C. Na + / K + ratio change map; Different lowercase letters in the figure under the same treatment conditions indicate significant differences between samples (P<0.05).

[0027] Figure 10 Proline and conductivity change map of wild-type and ProKCS11 :: VvKCS11- pCLB1301 overexpressed grape callus after 14 days of treatment with 0 mM and 100 mM NaCl; In the figure: A. Proline content change map; B. Relative conductivity change map; Different lowercase letters in the figure under the same treatment conditions indicate significant differences between samples (P<0.05). Specific implementation mode

[0028] The technical solutions of the present invention will be further explained below in combination with embodiments, but the protection scope of the present invention is not limited thereto.

[0029] The drugs, materials and reagents used in the examples are ordinary products on the market without special instructions. For the content not described in detail in the examples, they are all in accordance with the existing technologies in the art.

[0030] Example 1 Grape ProKCS11 Cloning of the promoter Using the genomic DNA of the leaves of 'Shiraz' grape as a template, the full-length promoter was cloned by nested PCR. ProKCS11 First, using KP-F-1 / KP-R-1 as primers, the target PCR product was amplified by PCR technology. Subsequently, using KP-F-2 / KP-R-2 as primers and the PCR product diluted 5-fold as a template, cloning amplification was carried out by PCR technology. The electrophoresis results showed that a promoter fragment with a length of about 1300 bp was obtained after amplification ( Figure 1 ), and the ProKCS11 promoter was obtained, and its nucleotide sequence is shown in SEQ ID NO.1.

[0031] Primer sequences: KP-F-1 is shown in SEQ ID NO.3; KP-R-1 is shown in SEQ ID NO.4; KP-F-2 is shown in SEQ ID NO.5; KP-R-2 is shown in SEQ ID NO.6.

[0032] Example 2 Grape ProKCS11 Promoter and GUS Construction and identification of the fusion expression vector In order to further verify the activity and expression pattern of the promoter cloned in Example 1, the present invention selected a GUS reporter gene fusion vector (1300GN) suitable for promoter expression analysis. Combining with the characteristics of the multiple cloning sites on 1300GN, ProKCS11 restriction enzyme sites (existing at the end of the KP–F-2 primer) were introduced into the 5' end of the Sal I promoter, and Bam HI restriction enzyme sites (existing at the end of the KP-R-2 primer) were introduced into the 3' end, and protective bases were introduced at both ends of the two restriction enzyme sites. The obtained ProKCS11 promoter was subjected to terminal modification by PCR reaction.

[0033] The PCR product was ligated into the pEASY-Blunt vector (TransGen Biotech, pEASY®-Blunt Zero Cloning Kit Blunt Zero gene cloning kit, CB501-01) by blunt-end ligation. Referring to the product manual, pEASY-Blunt and ProKCS11Ligation to obtain pEASYBlunt- ProKCS11 Subsequently, the 1300GN vector containing the GUS gene (this vector is based on the general vector pCAMBIA1300, the 35S promoter region is deleted, and the reporter system is replaced with the GUS gene; this vector can be constructed by the laboratory itself or by a biological company), and pEASYBlunt- containing the promoter fragment ProKCS11 were Sal I, Bam HI double digested. The digested fragments were separated by agarose gel electrophoresis and recovered. The results are shown in Figure 2 as shown in a. Subsequently, the ProKCS11 with restriction sites was ligated to the 1300GN vector using T4 ligase. The constructed fusion expression vector was transformed into Escherichia coli Trans 5α competent cells (TransGen Biotech, Trans5α Chemically Competent Cell Trans5α Cloning Competent Cells, CD201-01). The bacterial solution was spread on an LB plate containing 50 μg / mL Kan to screen for positive clones. The recombinant vector plasmid in the positive clones was extracted, and the recombinant expression vector was transferred into Agrobacterium tumefaciens GV3101 (Vazyme Biotech, GV3101 Chemically Competent Cell, AC1001) chemically competent cells. Single colonies with normal growth status were randomly selected on an LB plate containing 50 μg / mL Kan, and PCR detection was performed using M13F as shown in SEQ ID NO.7 and KP-R-2 as shown in Figure 2 as shown in b. The electrophoresis results showed that the positive bacteria had an obvious band at the position of 1500 bp. The positive bacteria were inoculated into an LB liquid medium containing 50 μg·mL -1 Kan and 25 μg·mL -1 rifampicin for amplification culture, and then sent to the company for sequencing for further verification. Finally, Agrobacterium tumefaciens GV3101 successfully carrying ProKCS11 :: GUS-1300GN was obtained.

[0034] Example 3 The Agrobacterium tumefaciens GV3101 carrying ProKCS11 :: GUS-1300GN obtained in Example 2 was used to infect Arabidopsis thaliana. The Arabidopsis thaliana infected through the inflorescence was continuously cultured until it flowered and produced seeds, and then these seeds were collected and dried. The dry transgenic Arabidopsis thaliana T0 generation seeds were disinfected and sown on a 1 / 2 MS solid medium (Coolaber, MS medium basal salts, PM519) containing 25 μg / mL hygromycin for screening, as shown in Figure 3 . The leaves of the obtained T1 generation transgenic plants were cut, and the total DNA of the transgenic plants was extracted. The transgenic Arabidopsis thaliana was identified by PCR as shown in Figure 4as shown

[0035] Example 4 GUS staining analysis of transgenic Arabidopsis thaliana under different stress induction conditions The transgenic Arabidopsis thaliana lines screened in Example 3 were sown on 1 / 2 MS solid medium containing 25 μg / mL hygromycin. When the plants grew to the 8th day, they were transplanted into 1 / 2 MS medium with different stress treatments respectively. These components included 0 mM NaCl (control), 100 mM NaCl, 20 μM ABA, 180 mM D-mannitol, 20 mM LiCl, and were treated in an environment at 25 °C for 48 hours. In the above treatments, ABA characterized the hormone signal response upstream of NaCl, and mannitol and LiCl could respectively analyze the responses of plants to osmotic stress and ionic toxicity equivalent to 100 mM NaCl by single factor. After the treatment, GUS staining analysis was carried out on its seedlings ( Figure 5 ), and the staining situation was observed under a microscope.

[0036] The results showed that under non-stress conditions, only the root-shoot junction of the transgenic lines showed a faint blue color. After applying environmental stress, the blue color significantly deepened and showed tissue specificity. Among them, 100 mM NaCl significantly deepened the GUS staining degree of the seedlings, and obvious expression trends could be observed in both roots and leaves. Under ABA treatment, the GUS staining degree had a tendency to deepen, but was significantly lower than that of other treatment groups. Under the osmotic stress condition simulated by D-mannitol treatment, it could be seen that the GUS staining deepened, but the GUS distribution was mainly concentrated in the old leaves, and the root expression was significantly lower than that of the 100 mM NaCl treatment group. GUS distribution was also visible in the LiCl treatment group, but this staining was mainly concentrated in the young leaves, and the root staining was slight, but significantly lower than that of the 100 mM NaCl treatment group. The above results indicate that ProKCS11 is a salt stress-inducible promoter. Under salt stress, this promoter drives the expression of downstream genes, and the expression shows a certain tissue specificity. Salt stress induction ProKCS11 initiates high expression of downstream genes at the whole plant level. Salt stress equivalent osmotic stress induces high expression of downstream genes in roots and old leaves, while salt stress equivalent ionic toxicity induces high expression of downstream genes in young leaves.

[0037] It can be seen from the above experimental results that the promoter provided by the present invention ProKCS11 has the functions of responding to salt stress, drought stress, osmotic stress, ionic stress and abscisic acid.

[0038] Example 5 Using the cDNA of the root of 'Shiraz' grape seedlings as a template and KCS-F and KCS-R as primers, a 1545 bp VvKCS11Sequence, the nucleotide sequence is as shown in SEQ ID NO.2.

[0039] Primer sequences: KCS-F is as shown in SEQ ID NO.8; KCS-R is as shown in SEQ ID NO.9.

[0040] The cDNA sequence with Bam HI and Sal Ⅰ restriction enzyme sites at both ends ( VvKCS11 sequence) was ligated to pTOPO (Polymerase, M5HiPer One-minute pTOPO-TA / Blunt Simple Cloning, MF889-01) in a blunt-end manner to obtain pTOPO- VvKCS11 , which was transformed into Escherichia coli Trans 5α competent cells, and positive clones were screened on an LB plate containing kanamycin (50 mg / L).

[0041] The promoter sequence ProKCS11 was amplified using primers KP-F-3 and KP-R-3 to obtain the promoter sequence with Kpn I and Bam HI restriction enzyme sites at both ends ProKCS11 . The promoter sequence with Kpn I and Bam HI restriction enzyme sites at both ends ProKCS11 was ligated to the vector pCLB1301NH (after purchasing pCAMBIA1301-35S-MCS-35S-EGFP-Hyg and deleting the 35S promoter) to obtain the recombinant plasmid pCLB1301NH- ProKCS11 , which was transformed into Escherichia coli Trans 5α competent cells, and positive clones were screened by colony PCR.

[0042] Primer sequences: KP-F-3 is as shown in SEQ ID NO.10; KP-R-3 is as shown in SEQ ID NO.11.

[0043] pTOPO- VvKCS11 was double-digested with Bam HI and Sal Ⅰ and then ligated to the similarly double-digested plant expression vector pCLB1301NH- ProKCS11 . After ligation, it was transformed into Escherichia coli Trans 5α competent cells, and positive clones were screened on an LB plate containing kanamycin (50 mg / L) ProKCS11 :: VvKCS11- pCLB1301.

[0044] Positive clones were screened by colony PCR with primers KCS-F and M13R as shown in SEQ ID NO.12, and the screening results are shown in Figure 6。Send the successfully verified plasmid to Personal Biotechnology Co., Ltd. in Qingdao for sequencing. Transform the successfully constructed plant expression vector into Agrobacterium tumefaciens competent cell GV3101 (Vidi Biotechnology, GV3101 Chemically Competent Cell, AC1001). Agrobacterium colony PCR detection is shown in Figure 6 。

[0045] Example 6 Suspend and culture the GV3101 Agrobacterium carrying ProKCS11 :: VvKCS11- pCLB1301 in LB medium, and adjust the OD600 value of the bacterial solution to 0.4 - 0.5. Incubate the adjusted bacterial solution at 28 °C and 180 - 200 rpm for about 2 hours. After the incubation, use the bacterial solution to infect the callus of 'Shiraz' grapes. Immerse the grape callus in the adjusted Agrobacterium bacterial solution and infect at 24 °C and 120 rpm for 15 min. After the infection, pour the mixture of the callus and the bacterial solution onto a mesh cloth and filter the mixture. After the filtration, scrape the callus onto filter paper and blot dry the bacterial solution. Subsequently, place the callus on GC medium (the composition is shown in Table 1, and all drugs are purchased from Coolaber Biotechnology Co., Ltd.) supplemented with 100 μM acetosyringone (AS) and co-culture in the dark for 48 hours. After the incubation, rinse the callus with sterile water containing 1000 mg / L timentin (Tmt), and transfer it to GC medium containing 1000 mg / L Tmt and 2.5 mg / L Hyg (hygromycin). Transfer once every 4 weeks until resistant callus grows. Observe the resistant callus under a fluorescence microscope. Obvious green fluorescence signals can be seen in the positive callus, as shown in Figure 7 , indicating that transgenic callus has been successfully obtained.

[0046] Table 1 Composition of GC medium

[0047] Example 7 ProKCS11 :: VvKCS11- Effect of overexpression of pCLB1301 gene on the growth and salt tolerance indexes of grape callus under salt stress Inoculate wild-type 'Shiraz' and ProKCS11 :: VvKCS11- pCLB1301 overexpressing callus in GC medium containing 0 mM and 100 mM NaCl, and incubate in the dark at 25 °C for 14 days. Observe the growth phenotype of the callus after 14 days as shown in Figure 8. As can be seen in the figure, after salt treatment, the growth of calli of both genotypes of grapes was inhibited to some extent, but the growth of transgenic calli was significantly better than that of the control group, specifically manifested by a lower growth inhibition effect after salt treatment.

[0048] Example 8 ProKCS11 :: VvKCS11- Overexpression of the pCLB1301 gene on the Na + and K + contents in grape calli under salt stress In plant cells, maintaining a low Na + concentration and a high K + concentration is a necessary prerequisite for various physiological processes, and salt stress will disrupt this balance. Maintaining ion homeostasis under salt stress is an important indicator for evaluating plant salt tolerance.

[0049] For wild-type and transgenic calli under different treatments, 0.1 g of each sample was weighed into a 10 mL centrifuge tube, and 4 replicates were set for each strain's control and treatment. 1 mL of concentrated nitric acid was added to each centrifuge tube for nitrification overnight. After nitrification, 3 mL of ultrapure water was continued to be added and heated in a water bath at 95 °C for 4 - 5 h. After the liquid cooled, it was fixed to 10 mL. Coarse filtration was carried out with a 7 cm diameter filter paper, and then filtered through a 0.45 μm filter membrane and stored in a -20 °C refrigerator. The Na + , K + contents of the samples were measured using a flame spectrophotometer, and the results are shown in Figure 9 . It can be seen that under salt stress, both wild-type and transgenic grape calli showed a trend of increasing Na + and decreasing K + , but this trend was more significant in wild-type calli. It indicates that ProKCS11 promoter-induced VvKCS11 overexpression can inhibit the accumulation of Na + in cells and promote the absorption of K + , maintaining the intracellular Na + , K + ion balance.

[0050] Example 9 Proline, as an osmotic adjustment substance, has a positive correlation between its accumulation amount and the ability to resist osmotic stress. After drawing the proline standard curve, the proline content in the sample was measured. About 0.1 g of callus was weighed and the weight was recorded. The tube wall was rinsed several times with 1.5 mL of 3% sulfosalicylic acid, and the callus and sulfosalicylic acid were transferred to a 2 mL Ep tube and placed in a boiling water bath for 10 min. After cooling in an ice bath, it was centrifuged at 12000 rpm for 10 minutes. 1 mL of the supernatant was taken into a 10 mL centrifuge tube, 1 mL of glacial acetic acid and 1.5 mL of ninhydrin color-developing solution were added, and after boiling in a water bath for 40 min and then cooling in an ice bath, 2.5 mL of toluene was added to oscillate and extract the red substance. After stratification, 200 μL of the toluene layer was taken, and using toluene as the blank control, the absorbance was measured at 520 nm with an enzyme-linked immunosorbent assay (ELISA) reader. The calculation method of proline is as follows: Proline (μg / g FW) =

[0051] In the calculation formula, C is the proline content in the extract (μg, obtained from the standard curve), V1 is the total volume of the extract (1.5 mL), V2 is the volume taken for determination (0.2 mL), and W is the sample weight (g).

[0052] The results of proline determination are shown in Figure 10 A. Salt stress induced ProKCS11 :: VvKCS11- the accumulation of proline in pCLB1301 transgenic grape callus, while no significant change in proline was observed in wild-type grape callus. This indicates that transgenic grape callus has better osmotic adjustment ability under salt stress.

[0053] Relative electrical conductivity characterizes the selectivity and stability of plant cell membranes. 0.2 g of grape callus was taken and placed in a test tube containing 4 mL of distilled water, and shaken in a constant temperature shaker at 28 °C and 120 rpm for 4 hours. The initial electrical conductivity of the callus (denoted as L1) and the electrical conductivity of the distilled water (denoted as L0) were measured respectively using a P902 type conductivity meter. Subsequently, the test tube was placed in boiling water and heated for about half an hour, taken out and cooled to room temperature, shaken to make the liquid uniform, and after standing for 10 minutes, the final electrical conductivity of the callus (denoted as L2) was measured. The relative electrical conductivity (REL) of the callus was calculated according to the following formula: REL(%) = (L1 - L0) × 100 / (L2 - L0).

[0054] The results of relative electrical conductivity determination are shown in Figure 10 B. Under salt stress, the electrical conductivity of wild-type grape callus increased significantly, indicating that salt stress caused damage to the cell membrane, reduced the selective permeability of the cell membrane, and caused the outflow of cell contents. While in ProKCS11 :: VvKCS11-In the pCLB1301 transgenic grape callus, the increase in relative electrical conductivity was significantly less than that of the wild type, indicating that the transgenic grape callus could maintain the stability of the cell membrane to a certain extent under salt stress.

[0055] In summary, the promoter provided by the present invention ProKCS11 has the functions of responding to salt stress, drought stress, osmotic stress, ionic stress and abscisic acid. Salt stress, osmotic stress equivalent to the salt stress effect, and ionic toxicity equivalent to the salt stress effect can all induce the high expression of the gene driven by this promoter. For example, by using this promoter to drive VvKCS11 overexpression can enhance the salt tolerance of transgenic grape callus, improve the salt tolerance of plants, and can be used to cultivate salt-tolerant transgenic plants; the promoter provided by the present invention can be used ProKCS11 to cultivate transgenic plants with stress resistance.

Claims

1. A promoter responsive to salt stress, drought stress, osmotic stress, ionic stress or abscisic acid ProKCS11 , characterized in that The promoter ProKCS11 has a nucleotide sequence as shown in SEQ ID NO.

1.

2. A biological material, characterized in that, The biomaterial is a recombinant vector, expression cassette, recombinant cell or recombinant bacterium containing the promoter with the nucleotide sequence shown in SEQ ID NO.

1. ProKCS11 ​ 3. The promoter according to claim 1 ProKCS11 or the use of the biological material according to claim 2 in any of the following: ① Preparation of anti-transgenic plant breeding agents; ② Improvement of plant stress resistance; ③ Cultivation of anti-transgenic plants with stress resistance.

4. The application according to claim 3, wherein The stress resistance mentioned above is salt stress resistance, drought stress resistance, osmotic stress resistance or ion stress resistance.

5. The application according to claim 3, wherein Expressing a target gene in a plant by a promoter ProKCS11 ​ 6. The application according to claim 5, characterized in that, The target gene mentioned above is a stress resistance gene.

7. The application according to claim 6, characterized in that The stress resistance genes include salt tolerance genes and drought resistance genes.

8. The application according to claim 7, wherein The stress resistance gene is VvKCS11 gene.

9. The application according to claim 8, wherein The VvKCS11 nucleotide sequence of the gene is shown in SEQ ID NO.

2.

10. The application according to claim 3, characterized in that The plant mentioned above is grape.

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