Grape VvKCS11 promoter and application thereof

Abstract

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

Description

Technical Field

[0001] This invention belongs to the field of plant genetic engineering technology, specifically relating to a grape... VvKCS11 Promoters and their applications. Background Technology

[0002] Soil salinization is a major problem threatening agricultural production and the ecological environment, seriously threatening crop yields and sustainable agricultural development. Grapes ( Vitis vinifera Grapes (L.) are an important economic fruit tree with high economic and nutritional value. Saline soils are distributed across many grape-producing areas in my country. Although grapes have a certain degree of salt tolerance, their growth is limited and yields decrease under high salinity conditions, severely hindering the sustainable development of my country's grape industry.

[0003] Under salt stress, the cork layer in grape roots thickens and occurs closer to the root tip, effectively preventing sodium absorption. + and Cl - Transported to the central column via apoplastic flow. β-Ketoacyl CoA synthase (KCS) participates in fatty acid chain elongation reactions and is the rate-limiting enzyme in the synthesis of suberin precursors. Studies have shown that grapes... VvKCS11 It is associated with the formation of the cork layer in the roots, and overexpression of this gene can significantly improve the salt tolerance of transgenic Arabidopsis thaliana.

[0004] CaMV35S Constitutive promoters, characterized by ease of operation and stable expression, have played a crucial role in plant stress resistance gene function research for many years. However, while stable expression of stress resistance genes throughout the entire plant can enhance plant resistance, it often leads to varying degrees of growth retardation, limiting their application in breeding. Differences in promoters of plant stress resistance-related genes are a significant reason for inter-varietal differences in resistance during plant evolution and domestication. These promoters are induced by salt stress, and their driven downstream genes often influence plant salt tolerance through tissue-specific expression. Existing research has identified… VvKCS11 Its role in plant salt tolerance, but verification of this function is achieved through constitutive promoters. CaMV35S The process was carried out, for VvKCS11 Research on promoters is still in its infancy; will this promoter affect... VvKCS11 Its salt tolerance effect in plants is still unclear. Summary of the Invention

[0005] To address the shortcomings of existing technologies, this invention provides a grape VvKCS11 Promoters and their applications.

[0006] The technical solution of the present invention is as follows:

[0007] A promoter that responds to salt stress, drought stress, osmotic stress, ion stress, or abscisic acid. ProKCS11 The promoter ProKCS11 The nucleotide sequence is shown in SEQ ID NO.1.

[0008] A biomaterial, said biomaterial being a nucleotide sequence containing the nucleotide sequence shown in SEQ ID NO.1. ProKCS11 Recombinant vectors, expression cassettes, recombinant cells, or recombinant bacteria that act as promoters.

[0009] The above promoter ProKCS11 Or the application of the above-mentioned biological materials in any of the following:

[0010] ① Preparation of plant breeding agents with anti-retroviral genes;

[0011] ② Improve plant stress resistance;

[0012] ③ Cultivate plants with anti-retroviral genes.

[0013] According to a preferred embodiment of the present invention, the stress resistance in the application is resistance to salt stress, drought stress, osmotic stress, or ion stress.

[0014] According to a preferred embodiment of the invention, the application is via promoter in plants. ProKCS11 To express the target gene.

[0015] More preferably, the target gene is a stress-resistance gene.

[0016] More preferably, the stress-resistance genes include salt-tolerant genes and drought-resistant genes.

[0017] More preferably, the stress-resistance gene is VvKCS11 Gene.

[0018] Further preferably, the VvKCS11 The nucleotide sequence of the gene is shown in SEQ ID NO.2.

[0019] According to a preferred embodiment of the invention, the plant is grape.

[0020] Beneficial effects

[0021] 1. The promoter provided by this invention ProKCS11 It exhibits responsiveness to salt stress, drought stress, osmotic stress, ion stress, or abscisic acid. Under salt stress conditions, the promoter... ProKCS11 It can enhance the expression of its driver genes. This induction effect can occur under osmotic stress and ion toxicity, which are equivalent to salt stress. The gene expression it drives has a certain tissue specificity.

[0022] 2. In this invention, the promoter ProKCS11 drive VvKCS11 The transgenic grape callus overexpressed showed better salt tolerance, and its growth rate, membrane stability, and ion homeostasis were all superior to those of the wild type. Attached Figure Description

[0023] Figure 1 For grapes VvKCS11 PCR amplification diagram of gene promoter;

[0024] In the diagram: Lanes 1-3: VvKCS11 promoter ProKCS11 Amplified fragment; M: DL2000 DNA Marker.

[0025] Figure 2 plant expression vector ProKCS11 :: Construction of GUS-1300GN and PCR verification of Agrobacterium colonies;

[0026] In the figure: (a) pBlunt- ProKCS11 and the 1300GN vector was double-digested with SalI and BamHI; (b) ProKCS11 ::GUS-1300GN Agrobacterium colony PCR; M: DL2000 DNA Marker; Lanes 1-2: pBlunt- ProKCS11 Double-digested fragments; Lane 3: 1300 GN vector SalI and BamHI double-digested fragments; Lanes 4-10: ProKCS11 :: GUS-1300GN Agrobacterium colony PCR positive fragment.

[0027] Figure 3 This is a screening diagram of transgenic Arabidopsis thaliana in 1 / 2 MS medium with hygromycin.

[0028] Figure 4 This is a PCR identification diagram of transgenic Arabidopsis thaliana.

[0029] In the figure: M: DL2000 DNA Marker; lanes 1-4: PCR products of 4 Arabidopsis thaliana plants that were positive for resistance screening.

[0030] Figure 5 GUS staining analysis of transgenic Arabidopsis seedlings after 48 hours of treatment with different stress conditions;

[0031] In the figure: the control group is the one without any stress treatment. ProKCS11 :: GUS-1300GN transgenic Arabidopsis thaliana plants.

[0032] Figure 6 plant expression vector ProKCS11 :: VvKCS11-Construction of pCLB1301 and PCR verification of Agrobacterium colonies;

[0033] In the figure: (a) Double enzyme digestion diagram of the vector and target fragment, lanes 1-2: ProKCS11-pEASY Blunt Vector restriction enzyme digestion diagram; lanes 3-4: VvKCS11-pCLB Vector restriction enzyme digestion diagram; (b) ProKCS11 :: VvKCS11- pCLB1301 Agrobacterium colony PCR, lanes 1-8 show the PCR results of 8 single colonies; M1: DL5000 DNA Marker; M2: DL2000 DNA Marker.

[0034] Figure 7 Image showing verification of callus tissue from genetically modified grapes;

[0035] In the diagram: A. Callus tissue of wild-type grapes under open threshing floor; B. Callus tissue of wild-type grapes under open threshing floor. ProKCS11 :: VvKCS11- A. Callus tissue of grapes overexpressing pCLB1301; B. Callus tissue of wild-type grapes under fluorescence excitation; C. Callus tissue of wild-type grapes under fluorescence excitation; D. Callus tissue of wild-type grapes under fluorescence excitation. ProKCS11 :: VvKCS11- Image of grape callus overexpressing pCLB1301.

[0036] Figure 8 After treatment with 0 mM and 100 mM NaCl for 14 days, wild-type and ProKCS11 :: VvKCS11- Phenotypic diagram of grape callus tissue overexpressing pCLB1301.

[0037] Figure 9 After treatment with 0 mM and 100 mM NaCl for 14 days, wild-type and ProKCS11 :: VvKCS11- pCLB1301 overexpression in grape callus: ion changes;

[0038] In the diagram: A. Na + Content variation graph; B. K + Content variation graph; C. Na + / K + Ratio variation graph; different lowercase letters in the graph under the same treatment conditions indicate that the differences between samples reached the significance level (P<0.05).

[0039] Figure 10 After treatment with 0 mM and 100 mM NaCl for 14 days, wild-type and ProKCS11 :: VvKCS11- Figure showing changes in proline and conductivity in grape callus overexpressed with pCLB1301;

[0040] In the figure: A. Changes in proline content; B. Changes in relative conductivity; Different lowercase letters under the same treatment conditions in the figure indicate that the differences between samples reached a significant level (P<0.05). Detailed Implementation

[0041] The technical solution of the present invention will be further explained and described below with reference to the embodiments, but the scope of protection of the present invention is not limited thereto.

[0042] Unless otherwise specified, the drugs, materials and reagents used in the examples are commercially available products. Any content not described in detail in the examples is based on the prior art.

[0043] Example 1

[0044] Grape ProKCS11 promoter cloning

[0045] Using genomic DNA from 'Shiraz' grape leaves as a template, nested PCR cloning was employed to obtain... ProKCS11 The full-length promoter was obtained. First, using KP-F-1 / KP-R-1 primers, PCR amplification was performed to obtain the PCR product containing the target. Then, using KP-F-2 / KP-R-2 primers, a 5-fold diluted PCR product was used as a template for cloning and amplification. Electrophoresis results showed that a promoter fragment of approximately 1300 bp was obtained after amplification. Figure 1 ),get ProKCS11 The promoter has a nucleotide sequence as shown in SEQ ID NO.1.

[0046] Primer sequences: KP-F-1 as shown in SEQ ID NO.3; KP-R-1 as shown in SEQ ID NO.4; KP-F-2 as shown in SEQ ID NO.5; KP-R-2 as shown in SEQ ID NO.6.

[0047] Example 2

[0048] Grape ProKCS11 promoter and GUS Construction and identification of fusion expression vectors

[0049] To further verify the activity and expression pattern of the promoter cloned in Example 1, this invention selected a promoter suitable for promoter expression analysis. GUS The reporter gene fusion vector (1300GN), combined with the characteristics of the multiple cloning site on 1300GN, can be used to... ProKCS11 5' end introduction of the promoter Sal I The restriction enzyme site (located at the end of the KP–F-2 primer) is introduced at the 3' end. Bam HIThe restriction enzyme sites (located at the ends of the KP-R-2 primers) are used, and protective bases are introduced at both ends of the two restriction enzyme sites. The resulting enzymes are then processed via PCR. ProKCS11 The promoter undergoes terminal modification.

[0050] The PCR product was ligated into the pEASY-Blunt vector (PEASY®-Blunt Zero Cloning Kit, CB501-01) using blunt-end ligation. The pEASY-Blunt vector was then ligated into the pEASY-Blunt vector using the product instructions. ProKCS11 The connection obtained pEASYBlunt- ProKCS11 Subsequently, the 1300GN vector containing the GUS gene (this vector is a modified version of the universal vector pCAMBIA1300, with the 35S promoter region deleted and the reporter system replaced by the GUS gene; this vector can be constructed by the laboratory itself or by a biotechnology company) and the pEASYBlunt vector containing the promoter fragment were tested. ProKCS11 conduct Sal I、Bam HI Double enzyme digestion. Agarose gel electrophoresis was used to separate and recover the digested fragments, as shown in the following results. Figure 2 As shown in Figure a. Subsequently, T4 ligase was used to ligate the enzyme-ligated molecules. ProKCS11 The fusion expression vector was ligated into a 1300GN vector. The constructed fusion expression vector was transformed into *E. coli* Trans 5α competent cells (Trans5α Chemically Competent Cell, CD201-01). The bacterial culture was plated on LB agar plates containing 50 μg / mL Kan to screen for positive clones. The recombinant vector plasmid was extracted from the positive clones and transformed into *Agrobacterium* GV3101 (Weidi Biotechnology, GV3101 Chemically Competent Cell, AC1001) chemically competent cells. Single colonies with normal growth were randomly selected from LB agar plates containing 50 μg / mL Kan, and PCR detection was performed using M13F (as shown in SEQ ID NO. 7) and KP-R-2. Figure 2 Electrophoresis results showed that bacteria with a distinct band at a size of 1500 bp were positive. The positive bacteria were inoculated into a solution containing 50 µg / mL... -1 Kan and 25 µg·mL -1 The culture was expanded in LB liquid medium containing rifampicin and then sent to the company for sequencing further validation. The virus was ultimately successfully carried out. ProKCS11 :: Agrobacterium GV3101 of GUS-1300GN.

[0051] Example 3

[0052] The carrier obtained in Example 2 ProKCS11 :: Agrobacterium GV3101 of GUS-1300GN was used to infect Arabidopsis thaliana. The inflorescence-infected Arabidopsis were continuously cultured until flowering and seed production, after which the seeds were collected and dried. The dried transgenic Arabidopsis T0 generation seeds were sterilized and sown on 1 / 2 MS solid medium (Coolaber, MS medium base salt, PM519) containing 25 μg / mL hygromycin for selection. Figure 3 As shown. Leaves of the obtained T1 generation transgenic plants were cut, and total DNA was extracted from the transgenic plants. The transgenic Arabidopsis thaliana was identified by PCR. Figure 4 As shown.

[0053] Example 4

[0054] GUS staining analysis of transgenic Arabidopsis under different stress induction conditions

[0055] The Arabidopsis transgenic lines screened in Example 3 were sown in 1 / 2 MS solid medium containing 25 μg / mL hygromycin. On day 8, the plants were transplanted into 1 / 2 MS medium containing different stress treatments, including 0 mM NaCl (control), 100 mM NaCl, 20 μM ABA, 180 mM D-mannitol, and 20 mM LiCl, and treated at 25°C for 48 hours. In these treatments, ABA characterized the upstream hormone signaling response to NaCl, while mannitol and LiCl were used for single-factor analysis of the plant's response to osmotic stress and ion toxicity equivalent to 100 mM NaCl. After treatment, GUS staining analysis was performed on the seedlings. Figure 5 ), and observe the staining under a microscope.

[0056] The results showed that under no stress conditions, only the root-stem junction of the transgenic lines showed a faint blue color, while under environmental stress, the blue color deepened significantly and showed tissue specificity. Specifically, 100 mM NaCl significantly deepened the GUS staining in seedlings, and a clear expression trend was observed in both roots and leaves. ABA treatment showed a tendency for GUS staining to deepen, but it was significantly lower than other treatment groups. Under D-mannitol treatment simulating osmotic stress, GUS staining deepened, but its distribution was mainly concentrated in older leaves, with root expression significantly lower than the 100 mM NaCl treatment group. GUS distribution was also observed in the LiCl treatment group, but the staining was mainly concentrated in young leaves, with slight root staining, but significantly lower than the 100 mM NaCl treatment group. These results indicate that... ProKCS11 This is a salt stress-inducible promoter; under salt stress, this promoter drives the expression of downstream genes, and the expression exhibits certain tissue specificity. Salt stress induces… ProKCS11The downstream genes were highly expressed throughout the plant. Salt stress, equivalent to osmotic stress, induced high expression of the downstream genes in the roots and older leaves, while salt stress, equivalent to ion toxicity, induced high expression of the downstream genes in young leaves.

[0057] The experimental results above show that the promoter provided by this invention... ProKCS11 It has the ability to respond to salt stress, drought stress, osmotic stress, ion stress and abscisic acid.

[0058] Example 5

[0059] Using 'Shiraz' grape seedling root cDNA as a template, a 1545 bp DNA sequence was cloned using KCS-F and KCS-R primers. VvKCS11 The sequence, the nucleotide sequence is shown in SEQ ID NO.2.

[0060] Primer sequences: KCS-F as shown in SEQ ID NO.8; KCS-R as shown in SEQ ID NO.9.

[0061] Bring both ends Bam HI and Sal Ⅰ cDNA sequence of restriction enzyme sites ( VvKCS11 The sequence is joined with pTOPO (aggregate beauty, M5HiPer One-minute pTOPO-TA / Blunt Simple Cloning, MF889-01) with blunt ends to obtain pTOPO- VvKCS11 The cells were transformed into Escherichia coli Trans 5α competent cells, and positive clones were screened on LB plates containing kanamycin (50 mg / L).

[0062] Amplification of promoter sequence using primers KP-F-3 and KP-R-3 ProKCS11 , to obtain both ends of the belt Kpn I and Bam HI promoter sequence of restriction enzyme sites ProKCS11 , bring both ends Kpn I and Bam HI promoter sequence of restriction enzyme sites ProKCS11 Ligating it with the pCLB1301NH vector (purchased pCAMBIA1301-35S-MCS-35S-EGFP-Hyg, then deleting the 35S promoter) yields pCLB1301NH- ProKCS11 The recombinant plasmid was transformed into E. coli Trans 5α competent cells, and positive clones were screened by colony PCR.

[0063] Primer sequences: KP-F-3 as shown in SEQ ID NO.10; KP-R-3 as shown in SEQ ID NO.11.

[0064] pTOPO- VvKCS11 use Bam HI and Sal Ⅰ After double digestion, it was compared with the plant expression vector pCLB1301NH- which was also double-digested. ProKCS11 After ligation, the cells were transformed into *E. coli* Trans 5α competent cells, and positive clones were selected on LB agar plates containing kanamycin (50 mg / L). ProKCS11 :: VvKCS11- pCLB1301.

[0065] Positive clones were screened by colony PCR using primers KCS-F and M13R as shown in SEQ ID NO.12. The screening results are shown below. Figure 6 The successfully validated plasmid was sent to Qingdao Paisenno Biotechnology Co., Ltd. for sequencing. The successfully constructed plant expression vector was transformed into Agrobacterium-competent GV3101 cells (Weidi Biotechnology, GV3101 Chemically Competent Cell, AC1001). Agrobacterium colony PCR detection is shown in [link to data]. Figure 6 .

[0066] Example 6

[0067] The carrier obtained in Example 5 ProKCS11 :: VvKCS11- Agrobacterium GV3101 (pCLB1301) was suspended in LB medium and the OD600 value was adjusted to 0.4-0.5. The prepared bacterial suspension was incubated at 28 °C with shaking at 180-200 rpm for approximately 2 hours. After incubation, the bacterial suspension was used to infect 'Shiraz' grape callus. The grape callus was immersed in the prepared Agrobacterium suspension and incubated at 24 °C with shaking at 120 rpm for 15 min. After infection, the mixture of callus and bacterial suspension was poured onto a mesh cloth and filtered. After filtration, the callus was scraped onto filter paper and the bacterial suspension was blotted dry. Subsequently, the callus was placed on GC medium supplemented with 100 μM acetosyringone (AS) (components shown in Table 1; all reagents were purchased from Coollab Biotechnology Co., Ltd.) and incubated in the dark for 48 hours. After culture, the callus was rinsed with sterile water containing 1000 mg / L tmentin (Tmt) and transferred to GC medium containing 1000 mg / L tmt and 2.5 mg / L hygromycin. Transferring was repeated every 4 weeks until resistant callus emerged. The resistant callus was observed under a fluorescence microscope; positive callus showed a clear green fluorescent signal. Figure 7 This indicates that transgenic callus tissue has been successfully obtained.

[0068] Table 1. Composition of GC Culture Medium

[0069]

[0070] Example 7

[0071] ProKCS11 :: VvKCS11- Effects of pCLB1301 gene overexpression on grape callus growth and salt tolerance indicators under salt stress

[0072] Wild-type 'Shiraz' and ProKCS11 :: VvKCS11- pCLB1301-overexpressing callus was inoculated into GC medium containing 0 mM and 100 mM NaCl and cultured in the dark at 25°C for 14 days. The callus growth phenotype was observed after 14 days. Figure 8 As shown in the figure, salt treatment inhibited the growth of callus tissue in both grape genotypes to some extent, but the growth of transgenic callus tissue was significantly better than that of the control group, specifically in that the growth inhibition effect was lower after salt treatment.

[0073] Example 8

[0074] ProKCS11 :: VvKCS11- Overexpression of the pCLB1301 gene inhibits Na2+ in grape callus under salt stress. + and K + Content effect

[0075] In plant cells, maintaining low Na + Concentration and high K + Concentration is a necessary prerequisite for various physiological processes, and salt stress can disrupt this balance. The maintenance of ion homeostasis under salt stress is an important indicator for assessing plant salt tolerance.

[0076] 0.1 g of each wild-type and transgenic callus sample was weighed into a 10 mL centrifuge tube, with four replicates for each control and treatment line. 1 mL of concentrated nitric acid was added to each centrifuge tube for nitrification overnight. After nitrification, 3 mL of ultrapure water was added, and the mixture was heated in a 95°C water bath for 4-5 h. After cooling, the volume was adjusted to 10 mL. The mixture was coarsely filtered through 7 cm diameter filter paper, then filtered through a 0.45 μm filter membrane and stored at -20°C. The Na content of the samples was determined using a flame spectrophotometer. + K + Content, the results are shown in Figure 9 It is evident that under salt stress, both wild-type and transgenic grape callus tissues exhibited Na+ deficiency. + Increase and K + The trend is decreasing, but this trend is more pronounced in wild-type callus. This indicates... ProKCS11 Promoter-induced VvKCS11Overexpression can inhibit Na + Accumulation within cells and promotion of K + Absorption, maintaining intracellular Na + K + Ion balance.

[0077] Example 9

[0078] Proline, as an osmotic regulator, shows a positive correlation between its accumulation and resistance to osmotic stress. After plotting a proline standard curve, the proline content in the samples was measured. Approximately 0.1 g of callus tissue was weighed and its weight recorded. The tube walls were rinsed several times with 1.5 mL of 3% sulfosalicylic acid. The callus tissue and sulfosalicylic acid were transferred to a 2 mL Eppendorf tube and incubated in a boiling water bath for 10 min. After cooling in an ice bath, the tube was centrifuged at 12000 rpm for 10 min. 1 mL of the supernatant was transferred to a 10 mL centrifuge tube, and 1 mL of glacial acetic acid and 1.5 mL of ninhydrin chromogenic solution were added. The tube was incubated in a boiling water bath for 40 min, then cooled in an ice bath. 2.5 mL of toluene was added and the mixture was shaken to extract the red substance. After separation, 200 μL of the toluene layer was taken, and the absorbance was measured at 520 nm using a microplate reader, with toluene as a blank control. The proline content was calculated as follows:

[0079] Proline (μg / g FW) =

[0080] 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 of the sample taken for measurement (0.2 mL), and W is the sample weight (g).

[0081] The results of the proline assay are shown below. Figure 10 A. Salt stress induced ProKCS11 :: VvKCS11- Proline accumulation was observed in pCLB1301 transgenic grape callus, while no significant changes in proline were observed in wild-type grape callus. This indicates that transgenic grape callus has better osmotic regulation under salt stress.

[0082] Relative conductivity characterizes the selectivity and stability of plant cell membranes. 0.2 g of grape callus tissue was placed in a test tube containing 4 mL of distilled water and shaken for 4 hours at 28℃ and 120 rpm in a constant-temperature shaker. The initial conductivity of the callus tissue (denoted as L1) and the conductivity of the distilled water (denoted as L0) were measured using a P902 conductivity meter. The test tube was then heated in boiling water for about half an hour, removed, cooled to room temperature, shaken to homogenize the liquid, and allowed to stand for 10 minutes before measuring the final conductivity of the callus tissue (denoted as L2). The relative conductivity (REL) of the callus tissue was calculated using the following formula:

[0083] REL(%)=(L1-L0)×100 / (L2-L0).

[0084] The results of the relative conductivity measurement are shown in Figure 10 In the medium B group, under salt stress, the electrical conductivity of wild-type grape callus tissue significantly increased, indicating that salt stress caused cell membrane damage, reduced cell membrane selective permeability, and led to the outflow of cell contents. Meanwhile... ProKCS11 :: VvKCS11- In pCLB1301 transgenic grape callus, the increase in relative conductivity was significantly smaller than that in wild type, indicating that transgenic grape callus can maintain cell membrane stability to a certain extent under salt stress.

[0085] In summary, the promoter provided by this invention ProKCS11 It exhibits responsiveness to salt stress, drought stress, osmotic stress, ion stress, and abscisic acid. Salt stress, osmotic stress (equivalent to salt stress), and ion toxicity (equivalent to salt stress) can all induce high expression of genes driven by this promoter. For example, using this promoter to drive... VvKCS11 Overexpression of this gene can enhance the salt tolerance of transgenic grape callus tissue, thereby improving plant salt tolerance and making it suitable for cultivating salt-tolerant transgenic plants. The promoter provided by this invention can be used... ProKCS11 Cultivate plants with anti-retroviral genes.

Claims

1. Promoter ProKCS11 or contains ProKCS11 Application of biomaterials with promoters in any of the following: ① Preparation of plant breeding agents with anti-retroviral genes; ② Improve plant stress resistance; ③ Cultivate plants with anti-retroviral genes; The promoter ProKCS11 The nucleotide sequence is shown in SEQ ID NO.1; The containing ProKCS11 Biomaterials containing promoters ProKCS11 Recombinant vectors, expression cassettes, recombinant cells, or recombinant bacteria for promoters; The resistance mentioned refers to resistance to salt stress; The application, in plants, via promoters ProKCS11 Express salt tolerance genes; The plants mentioned are grapes and Arabidopsis thaliana.

2. The application as described in claim 1, characterized in that, The gene is VvKCS11 Gene.

3. The application as described in claim 2, characterized in that, The VvKCS11 The nucleotide sequence of the gene is shown in SEQ ID NO.2.

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