Receptor-like kinase gene for improving broomrape parasitism resistance of sunflower and application of receptor-like kinase gene

Abstract

The invention relates to the technical field of plant genetic engineering, and provides a receptor-like kinase gene for improving Orobanche coerulescens parasitism resistance of sunflower and application of the receptor-like kinase gene. The invention discovers an amino acid sequence of the sunflower receptor kinase and a nucleotide sequence of the coding gene HaCERK1, and provides a target sequence for silencing the HaCERK1 gene, a virus-mediated gene silencing system and a silencing method based on the amino acid sequence, so that the expression level of the sunflower receptor kinase HaCERK1 gene can be effectively inhibited, and the expression level of the sunflower receptor kinase HaCERK1 gene can be effectively inhibited. And a virus-mediated gene silencing system can be systematically diffused in sunflower and sunflower orobanche tissues, so that the expression of the HaCERK1 gene is effectively interfered. According to the virus-mediated gene silencing system and method provided by the invention, the sunflower gene function research efficiency is remarkably improved, and the time and the cost are greatly reduced.

Description

Technical Field

[0001] The present invention relates to the technical field of plant genetic engineering. More specifically, the present invention relates to a receptor-like kinase gene for improving the resistance of sunflowers to Orobanche cumana parasitism and its application. Background Art

[0002] Sunflower is an important oil crop, commonly used in the production of vegetable oil and the processing of feed and industrial raw materials. Orobanche cumana Wallr is an annual root parasitic weed that mainly parasitizes on the roots of sunflowers. By establishing a parasitic relationship with the sunflower roots, it absorbs the nutrients and water of sunflowers to achieve complete parasitism. Once Orobanche cumana parasitizes on the roots of sunflowers, it will cause a decrease in the biomass and yield of sunflowers. In severe cases, the yield can be reduced by 80% or even result in a complete crop failure, and the oil content of sunflowers will also be seriously affected. Worldwide, the control of Orobanche cumana has always been a major problem.

[0003] Virus-induced gene silencing (VIGS) assay refers to a technical means in which when a plant is infected by a viral vector carrying a target gene fragment, it can induce the silencing of the target gene and show corresponding phenotypic changes. It has the characteristics of simple operation, rapid acquisition of phenotypes, independence from plant genetic transformation, avoidance of lethal mutations, research on the functions of complex redundant genes, and rapid comparison of gene functions between different species. Therefore, it has been widely used in some species with complex genetic backgrounds and difficult genetic transformation.

[0004] Since Orobanche cumana is a root parasitic weed, its underground growth stage has already had a serious impact on the growth of host plants. Therefore, the critical period for controlling Orobanche cumana is the underground growth stage, that is, the early infection stage of Orobanche cumana on sunflowers. At present, the measures for controlling Orobanche cumana mainly include biological control, spraying herbicides, manual removal, inducing eradication using trap crops, chemical control, and breeding of resistant species. However, each method has certain limitations to some extent. The key gene of the sunflower receptor-like kinase, silencing the expression of this gene can cause an albino phenotype in plants, and the phenotype is obvious and easy to observe, which is a very important gene for developing the VIGS silencing technology. The existing public literature 1 (Host sunflower-induced silencing of parasitism-related genes confers resistance to invading Orobanche cumana. (2021). Plant Physiology, 185(2), 424-440) studied the gene silencing technology for the parasitic weed Orobanche cumana. In the study, the method of inducing gene silencing using the Tobacco rattle virus (TRV) vector was used to systematically silence the target gene HaTubulin, and successful cross-species gene silencing was achieved in Orobanche cumana, weakening the parasitic ability and growth of Orobanche cumana, showing the potential of VIGS in the control of parasitic plants. However, the current research on sunflower gene function is still relatively lagging behind. Its genome ploidy is complex, and the sunflower receptor-like kinase and its coding sequence are not clear. Summary of the Invention

[0005] To overcome the above defects of the prior art, the present invention provides a receptor-like kinase gene for improving the resistance of sunflowers to Orobanche cumana parasitism. Using the virus-mediated gene silencing technology, by designing specific target sequences, constructing VIGS vectors, Agrobacterium and its infection solutions, the expression level of the HaCERK1 gene can be effectively inhibited, revealing the positive regulation of the HaCERK1 gene against Orobanche cumana parasitism, improving the efficiency of sunflower gene function research, and at the same time significantly saving research time and resource costs, providing strong technical support for the genetic improvement of sunflower resistance.

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

[0007] A receptor-like kinase gene for improving the resistance of sunflowers to Orobanche cumana parasitism, and the amino acid sequence of the sunflower receptor-like kinase is shown in SEQ ID NO.1.

[0008] As a further solution of the present invention, the nucleotide sequence of the HaCERK1 gene encoding the sunflower receptor-like kinase is shown in SEQ ID NO.2.

[0009] Application of a receptor-like kinase gene for improving sunflower resistance to Orobanche parasitism. By silencing the sunflower receptor-like kinase HaCERK1 gene, the parasitism of Orobanche weeds on sunflowers is enhanced, wherein the Orobanche weeds include Orobanche cumana Wallr.

[0010] As a further aspect of the present invention, the nucleotide sequence of the target sequence for silencing the sunflower receptor-like kinase HaCERK1 gene is as shown in SEQ ID NO.3.

[0011] As a further aspect of the present invention, the virus-mediated gene silencing vector for silencing the sunflower receptor-like kinase HaCERK1 gene includes the target sequence for silencing the sunflower receptor-like kinase HaCERK1 gene.

[0012] As a further aspect of the present invention, the backbone vector of the virus-mediated gene silencing vector for silencing the sunflower receptor-like kinase HaCERK1 gene includes the pTRV2 vector.

[0013] As a further aspect of the present invention, the Agrobacterium for silencing the sunflower receptor-like kinase HaCERK1 gene includes the virus-mediated gene silencing vector for silencing the sunflower receptor-like kinase HaCERK1 gene.

[0014] As a further aspect of the present invention, the infiltration solution for silencing the sunflower receptor-like kinase HaCERK1 gene includes the Agrobacterium for silencing the sunflower receptor-like kinase HaCERK1 gene.

[0015] As a further aspect of the present invention, the absorbance value of the Agrobacterium for silencing the sunflower receptor-like kinase HaCERK1 gene in the infiltration solution for silencing the sunflower receptor-like kinase HaCERK1 gene at a wavelength of 600 nm is 0.6 - 1.0.

[0016] As a further aspect of the present invention, silencing the sunflower receptor-like kinase HaCERK1 gene includes the following steps:

[0017] Step S1, select healthy and plump sunflower seeds with a germination rate above 95% and remove their hulls.

[0018] Step S2, put the shelled sunflower seeds into distilled water and soak them at room temperature for 2 hours.

[0019] Step S3, after soaking, remove the seed coat.

[0020] Step S4, use a sterile needle to scratch the surface of each seed, and control the scratch within the seed epidermal layer without penetrating the internal tissue of the seed embryo.

[0021] Step S5: Prepare an infiltration solution for silencing the sunflower receptor-like kinase HaCERK1 gene. Before use, mix it evenly with pTRV1 at a volume ratio of 1:1, and measure its absorbance at a wavelength of 600 nm to ensure it is between 0.6 and 1.0.

[0022] Step S6: Completely immerse the scratched seeds in step S4 in the prepared infiltration solution, and soak them for more than 6 hours under dark conditions at room temperature.

[0023] Step S7: Take out the soaked seeds and wash them with distilled water at least three times, replacing the clean distilled water each time.

[0024] Step S8: Transfer the washed seeds to MS liquid medium and culture them for 1 day under dark conditions at room temperature.

[0025] Step S9: Transfer the cultured seeds into the pre-prepared soil mixed with Orobanche.

[0026] Compared with the prior art, the beneficial effects of the receptor-like kinase gene for improving sunflower resistance to Orobanche parasitism and its application in the present invention are as follows:

[0027] In the present invention, by using virus-induced gene silencing technology (VIGS) to specifically silence the sunflower receptor-like kinase HaCERK1 gene, effective inhibition of gene expression in sunflowers and the parasitic weed Orobanche has been successfully achieved, revealing the key mechanism of action of the HaCERK1 gene as a positive regulator of resistance to Orobanche parasitism. Experiments show that silencing the HaCERK1 gene significantly enhances the parasitic ability of Orobanche on sunflowers, further demonstrating the importance of this gene in the sunflower anti-parasitic network. The present invention improves the efficiency of sunflower gene function research, and at the same time significantly saves research time and resource costs, providing strong technical support for sunflower resistance genetic improvement. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 It is a construction diagram of the VIGS virus vector of the HaCERK1 gene of the present invention.

[0029] Figure 2 It is a comparison diagram of the enhanced effect of HaCERK1 gene silencing on Orobanche infection of sunflower roots in the present invention.

[0030] Figure 3 It is an impact diagram of HaCERK1 gene silencing on the sunflower leaf phenotype in the present invention.

[0031] Figure 4 It is a comparison diagram of the impact of HaCERK1 gene silencing on the overall above-ground growth of sunflower plants in the present invention.

[0032] Figure 5 Statistical chart of the plant height of sunflower plants in different treatment groups.

[0033] Figure 6 Comparison diagram of sunflower root tissue sections.

[0034] Figure 7 Electrophoresis diagram for detecting the expression of the HaCERK1 gene silencing of the present invention in Orobanche cumana tissue and sunflower roots.

[0035] Figure 8 Statistical analysis diagram of the effect of HaCERK1 gene silencing on the number and weight of Orobanche cumana parasitism.

[0036] Figure 9 Analysis diagram of the relative expression level of the HaCERK1 gene in sunflower roots and Orobanche cumana tissue.

[0037] In the figure, pTRV: pTRV vector, control vector, a virus-mediated gene silencing vector that does not contain any target sequence; pTRV2-HaCERK1: pTRV2-HaCERK1 vector, a virus-mediated gene silencing vector containing the target sequence of the HaCERK1 gene, used to silence the HaCERK1 gene; OD600 = 0.8: the absorbance value at a wavelength of 600 nm is 0.8; Plant height(cm): plant height (cm); Oc: Orobanche cumana, sunflower broomrape; Vs: vascular system of the sunflower root, the vascular system of the sunflower root; pTRV-HaCERK1-1: the first experiment of pTRV-HaCERK1; pTRV-HaCERK1-2: the second experiment of pTRV-HaCERK1; pTRV-HaCERK1-3: the third experiment of pTRV-HaCERK1; Control: untreated group; CK: control group; M: Marker, molecular weight standard; mark: marker. Detailed implementation manners

[0038] Next, the technical solutions in this embodiment will be clearly and completely described in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without making creative efforts shall fall within the protection scope of the present invention.

[0039] A receptor-like kinase gene for improving the resistance of sunflowers to Orobanche cumana parasitism, the gene is the HaCERK1 gene, and the amino acid sequence of the sunflower receptor-like kinase is shown in SEQ ID NO.1, specifically as follows:

[0040]

[0041]

[0042]

[0043]

[0044]

[0045] The nucleotide sequence of the HaCERK1 gene encoding the sunflower receptor-like kinase is shown in SEQ ID NO.2, as follows:

[0046]

[0047]

[0048]

[0049]

[0050] An application of a receptor-like kinase gene for improving sunflower resistance to Orobanche parasitism. By silencing the sunflower receptor-like kinase HaCERK1 gene, the parasitism rate of sunflower parasitic weeds on sunflowers is enhanced, wherein the sunflower parasitic weeds include Orobanche cumana Wallr.

[0051] The nucleotide sequence of the target sequence for silencing the sunflower receptor-like kinase HaCERK1 gene is shown in SEQ ID NO.3, as follows:

[0052]

[0053]

[0054] The virus-mediated gene silencing vector for silencing the sunflower receptor-like kinase HaCERK1 gene includes the target sequence for silencing the sunflower receptor-like kinase HaCERK1 gene.

[0055] The backbone vector of the virus-mediated gene silencing vector for silencing the sunflower receptor-like kinase HaCERK1 gene includes the pTRV2 vector.

[0056] The Agrobacterium for silencing the sunflower receptor-like kinase HaCERK1 gene includes the virus-mediated gene silencing vector for silencing the sunflower receptor-like kinase HaCERK1 gene.

[0057] The infiltration solution for silencing the sunflower receptor-like kinase HaCERK1 gene includes the Agrobacterium for silencing the sunflower receptor-like kinase HaCERK1 gene.

[0058] In the infiltration solution for silencing the sunflower receptor-like kinase HaCERK1 gene, the absorbance value of the Agrobacterium for silencing the sunflower receptor-like kinase HaCERK1 gene at a wavelength of 600 nm is 0.6 - 1.0.

[0059] Silencing the sunflower receptor-like kinase HaCERK1 gene includes the following steps:

[0060] Step S1: Select healthy and plump sunflower seeds with a germination rate above 95% and remove their shells.

[0061] Step S2: Put the shelled sunflower seeds into distilled water and soak them at room temperature for 2 hours.

[0062] Step S3: After soaking, remove the seed coats.

[0063] Step S4: Use a sterile needle to scratch the surface of each seed, and control the scratch within the seed epidermal layer without penetrating the internal tissue of the embryo, so that the embryo is intact and conducive to subsequent infection treatment.

[0064] Step S5: Prepare the infiltration solution for silencing the sunflower receptor-like kinase HaCERK1 gene, mix it evenly with pTRV1 in a 1:1 equal volume before use, and measure its absorbance value at a wavelength of 600 nm to ensure it is between 0.6 and 1.0.

[0065] Step S6: Completely immerse the seeds after the scratch treatment in step S4 in the prepared infiltration solution, soak them in the dark at room temperature for more than 6 hours, and gently stir the infiltration solution appropriately during this period to ensure full contact between the infiltration solution and the seed surface.

[0066] Step S7: Take out the soaked seeds and wash them with distilled water at least three times, replacing the clean distilled water each time, to remove the excess infection solution attached to the seed surface and avoid affecting subsequent cultivation.

[0067] Step S8: Transfer the washed seeds to MS liquid medium and culture them in the dark at room temperature for 1 day.

[0068] Step S9: Transfer the cultured seeds into the pre-prepared soil mixed with Orobanche, gently cover with fine soil to ensure full contact between the seeds and the soil, and regularly spray an appropriate amount of water to keep the soil moist to promote seed germination and subsequent growth.

[0069] Based on the above advantages, the target sequence provided by the present invention can effectively reduce the expression level of the sunflower HaCERK1 gene. Compared with the empty vector control, the relative expression level of the HaCERK1 gene decreased by 54.19% - 66.93%.

[0070] Example 1

[0071] The virus-mediated gene silencing vector for silencing the receptor-like kinase HaCERK1 gene in sunflower in the embodiments of the present invention includes a target sequence; wherein, the backbone vector of the virus-mediated gene silencing vector is preferably a pTRV2 vector, and the target sequence is preferably inserted between the BamHⅠ and KpnⅠ restriction enzyme sites of the pTRV2 vector.

[0072] Example 2

[0073] The Agrobacterium for silencing the receptor-like kinase HaCERK1 gene in sunflower in the embodiments of the present invention includes the virus-mediated gene silencing vector; wherein, the Agrobacterium is preferably Agrobacterium, more preferably Agrobacterium tumefaciens, and even more preferably Agrobacterium tumefaciens GV3101.

[0074] Example 3

[0075] The infection solution for silencing the receptor-like kinase HaCERK1 gene in sunflower in the embodiments of the present invention includes the Agrobacterium; wherein, the OD600 value of the Agrobacterium is preferably 0.6 to 1.0, more preferably 0.8; the infection solution further includes Agrobacterium containing an auxiliary vector and an infection medium; the Agrobacterium containing the auxiliary vector is preferably Agrobacterium, more preferably Agrobacterium tumefaciens, and even more preferably Agrobacterium tumefaciens GV3101; the auxiliary vector is preferably a pTRV1 vector; the infection medium is preferably based on MS liquid medium, and further preferably includes 10 mM magnesium chloride, 10 mM 2-morpholinoethanesulfonic acid, and 200 μM acetosyringone.

[0076] Example 4

[0077] The method for silencing the receptor-like kinase HaCERK1 gene in sunflower in the embodiments of the present invention includes the following steps:

[0078] Step S1, select healthy and plump sunflower seeds with a germination rate of over 95% and remove their husks;

[0079] Step S2, put the shelled sunflower seeds into distilled water and soak them at room temperature for 2 hours;

[0080] Step S3, after soaking, remove the seed coat;

[0081] Step S4: Use a sterile needle to make a wound on the surface of each seed, and control the scratch within the seed epidermal layer without penetrating the internal tissue of the embryo;

[0082] Step S5: Prepare an infiltration solution for silencing the receptor-like kinase HaCERK1 gene in sunflower. Before use, mix it evenly with pTRV1 at a volume ratio of 1:1, and measure its absorbance at a wavelength of 600 nm to ensure it is between 0.6 and 1.0;

[0083] Step S6: Completely immerse the seeds treated by scratching in step S4 in the prepared infiltration solution, and soak for more than 6 hours under dark conditions at room temperature;

[0084] Step S7: Take out the soaked seeds and wash them with distilled water at least three times, and change the distilled water each time;

[0085] Step S8: Transfer the washed seeds to MS liquid medium and culture them for 1 day under dark conditions at room temperature;

[0086] Step S9: Transfer the cultured seeds into the pre-prepared soil mixed with Orobanche.

[0087] Example 5

[0088] The phenotypic statistics after the treatment of the receptor-like kinase gene that improves the resistance of sunflower to Orobanche parasitism in the present invention include: as Figure 2 shown in the comparison diagram of the enhanced effect of HaCERK1 gene silencing on Orobanche infection in the roots of sunflower in the present invention, which shows the Orobanche parasitism status underground of sunflower under different treatments. The number of Orobanche in the pTRV2-HaCERK1 treatment group is significantly more than that in the control group, and the damage is more serious; as Figure 3 shown in the diagram of the effect of HaCERK1 gene silencing on the leaf phenotype of sunflower in the present invention, which shows the morphological changes of the above-ground leaves of sunflower under different treatments. The leaves of pTRV-HaCERK1 have obvious shrinkage, notches and albino spots under VIGS treatment; as Figure 4 shown in the comparison diagram of the effect of HaCERK1 gene silencing on the overall growth of the above-ground part of sunflower plants in the present invention, which shows the comparison of the growth status of the above-ground plants between the control group and the pTRV2-HaCERK1 treatment group, Figure 5 shown in the statistical chart of the plant height of sunflower plants in different treatment groups. After silencing the HaCERK1 gene by VIGS, the sunflower plants are significantly shorter and the growth of the leaves becomes worse, which may be related to the serious Orobanche parasitism status underground affecting the normal growth of the plants.

[0089] Example 6

[0090] As Figure 6The figure shows a comparison diagram of sunflower root tissue sections. They are the tissue structures of the control group (pTRV2) and the group treated with the silenced HaCERK1 gene (pTRV-HaCERK1). The control group represents the sunflower root tissue without silencing the HaCERK1 gene, with a complete and tightly arranged cell structure, showing a normal cell tissue morphology; the group treated with the silenced HaCERK1 gene represents the root tissue after silencing the HaCERK1 gene through the pTRV-HaCERK1 vector. It can be observed that the cell structure of this tissue is loose and disordered, showing changes in the cell structure. This change indicates that after silencing the HaCERK1 gene, the resistance of sunflower roots is weakened, the cell structure is damaged, and further leads to an increase in the infestation of the parasitic weed Orobanche cumana Wallr. on sunflowers.

[0091] Example 7

[0092] As Figure 7 Shown is the electrophoresis diagram of the expression detection of the silenced HaCERK1 gene of the present invention in Orobanche cumana Wallr. tissue and sunflower roots, which is used to evaluate the effect of gene silencing treatment in different samples. The figure includes samples of Orobanche cumana Wallr. tissue and sunflower roots, and the expression of the HaCERK1 gene in each sample is detected by electrophoresis. In the samples of Orobanche cumana Wallr. tissue and sunflower roots, the pTRV-HaCERK1 treatment was carried out respectively to detect the silencing effect of this gene. The electrophoresis band conditions of different samples can be seen in the figure, including the treatment groups of 09, 010, 011, 012 of Orobanche cumana Wallr. tissue and H6, H7, H8, and H9 of sunflower roots. All samples show specific electrophoresis bands, indicating the expression of the gene after gene silencing treatment. By comparing the brightness and intensity of the bands, the expression level of the HaCERK1 gene in each sample can be evaluated.

[0093] Example 8

[0094] As Figure 8 Shown is the statistical analysis diagram of the effect of silencing the HaCERK1 gene on the number and weight of Orobanche cumana Wallr. parasitism. The ordinate in the figure represents the number and weight of Orobanche cumana Wallr. parasitism respectively. By comparing the untreated group, the control group (pTRV), and the gene silencing treatment group (pTRV-HaCERK1), the effect of gene silencing treatment on the number and weight of Orobanche cumana Wallr. parasitism can be observed. The figure shows the distribution of the number and weight of Orobanche cumana Wallr. parasitism in each group. Generally speaking, the number and weight of Orobanche cumana Wallr. parasitism in the pTRV-HaCERK1 treatment group are higher than those in the untreated group and the pTRV control group, indicating that silencing the HaCERK1 gene may lead to an increase in the number and weight of Orobanche cumana Wallr. parasitism, suggesting that the HaCERK1 gene may play an important role in resisting Orobanche cumana Wallr. parasitism.

[0095] Example 9

[0096] As Figure 9The figure shows the analysis of the relative expression levels of the HaCERK1 gene in sunflower roots and Orobanche aegyptiaca tissues. The bar graph in the figure shows the relative expression levels of the HaCERK1 gene in sunflower roots and Orobanche aegyptiaca tissues in different treatment groups. By comparing the gene expression between the pTRV control group and each pTRV-HaCERK1 treatment group, it can be seen that the gene silencing treatment significantly reduced the expression level of the HaCERK1 gene. The asterisks indicate statistical significance, showing that there are significant or highly significant differences between the gene silencing treatment group and the pTRV control group.

[0097] As described above, this is only the specific implementation manner of the present application, but the protection scope of the present application is not limited thereto. Any person skilled in the art within the technical scope disclosed by the present application can easily think of changes or substitutions, which should all be covered within the protection scope of the present application. Therefore, the protection scope of the present application should be subject to the protection scope of the claims.

[0098] Finally: The above are only the preferred embodiments of the present invention and are not used to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. A receptor kinase gene for improving sunflower resistance to Orobanche parasitism, characterized in that: The amino acid sequence of the sunflower receptor kinase is shown in SEQ ID NO.

1.

2. The receptor-like kinase gene for improving sunflower resistance to Orobanche parasitism according to claim 1, characterized in that: The nucleotide sequence of the HaCERK1 gene encoding the sunflower receptor kinase is shown in SEQ ID NO.

2.

3. An application of a receptor kinase-like gene for improving sunflower resistance to Orobanche parasitism, characterized in that: By silencing the sunflower receptor kinase HaCERK1 gene, the parasitism of sunflower parasitic weeds on sunflower is enhanced, wherein the sunflower parasitic weeds include sunflower broomrape.

4. The use of a receptor kinase-like gene for improving sunflower resistance to Orobanche parasitism according to claim 3, characterized in that: The nucleotide sequence of the target sequence for silencing the sunflower receptor kinase HaCERK1 gene is shown in SEQ ID NO.

3.

5. The use of a receptor kinase-like gene for improving sunflower resistance to Orobanche parasitism according to claim 3 or 4, characterized in that: The virus-mediated gene silencing vector for silencing the sunflower receptor kinase HaCERK1 gene comprises the target sequence for silencing the sunflower receptor kinase HaCERK1 gene.

6. The use of a receptor kinase-like gene for improving sunflower resistance to Orobanche parasitism according to claim 3, characterized in that: The backbone vector of the virus-mediated gene silencing vector used for silencing the sunflower receptor kinase HaCERK1 gene includes a pTRV2 vector.

7. The use of a receptor kinase-like gene for improving sunflower resistance to Orobanche parasitism according to claim 3 or 6, characterized in that: The Agrobacterium used for silencing the sunflower-like receptor kinase HaCERK1 gene includes the virus-mediated gene silencing vector for silencing the sunflower-like receptor kinase HaCERK1 gene.

8. The use of a receptor kinase-like gene for improving sunflower resistance to Orobanche parasitism according to claim 7, characterized in that: The infection solution for silencing the sunflower-like receptor kinase HaCERK1 gene includes the Agrobacterium for silencing the sunflower-like receptor kinase HaCERK1 gene.

9. The use of a receptor kinase-like gene for improving sunflower resistance to Orobanche parasitism according to claim 3 or 8, characterized in that: The absorbance value of the Agrobacterium used to silence the sunflower receptor kinase HaCERK1 gene in the infection solution used to silence the sunflower receptor kinase HaCERK1 gene is 0.6-1.0 at a wavelength of 600nm.

10. The use of a receptor kinase-like gene for improving sunflower resistance to Orobanche parasitism according to claim 3, characterized in that: Silencing the sunflower receptor kinase HaCERK1 gene comprises the following steps: Step S1, selecting healthy and full sunflower seeds with a germination rate of more than 95%, and shelling them; Step S2, placing the shelled sunflower seeds into distilled water and soaking them at room temperature for 2 hours; Step S3, after soaking is completed, peeling off the seed coat; Step S4, using a sterile needle to make a wound on the surface of each seed, and the wound is controlled within the seed epidermis without penetrating the internal tissue of the embryo; Step S5, preparing an impregnation solution for silencing the sunflower receptor kinase HaCERK1 gene, uniformly mixing it with an equal volume of pTRV1 at a ratio of 1:1 before use, and measuring its absorbance at a wavelength of 600 nm to ensure that it is between 0.6 and 1.0; Step S6, completely immersing the seeds that have been scratched in step S4 in the prepared impregnation solution, and soaking them in the dark at room temperature for more than 6 hours; Step S7, taking out the soaked seeds and washing them with distilled water at least three times, and changing the distilled water each time; Step S8, transferring the washed seeds to MS liquid culture medium and culturing them at room temperature in the dark for 1 day; Step S9, the cultured seeds are transferred into the pre-prepared soil mixed with Olabaster.

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