Signal protein RH1 for promoting phosphorus absorption and growth of plants and application of signal protein RH1

Abstract

The invention relates to a signal protein RH1 for promoting phosphorus absorption and growth of plants and application of the signal protein RH1, the amino acid sequence of the signal protein RH1 is shown as SEQ ID NO.1, and the plants include but are not limited to arabidopsis thaliana and oilseed rape.

Description

Technical Field

[0001] This invention belongs to the field of genetic engineering technology, specifically relating to a signaling protein RH1 that promotes phosphorus absorption and growth in plants and its applications. Background Technology

[0002] Fertilizer is an indispensable agricultural input for ensuring national food security, playing a vital role in improving crop yield and quality. my country is the world's largest consumer of fertilizers, but the overall efficiency of fertilizer application still needs improvement. In agricultural production, the excessive use of chemical fertilizers not only increases agricultural production costs and reduces the quality of agricultural products, but also causes prominent problems such as soil structure damage, decreased arable land productivity, and eutrophication of surface water, seriously hindering the development of modern agriculture and ecological environment construction. Therefore, developing and using green and efficient new fertilizers, and accelerating the reduction and efficiency improvement of fertilizer use, has become an inevitable choice for ensuring national food security and sustainable agricultural development.

[0003] In recent years, the research and application of immune signaling proteins have brought new breakthroughs to the development of green fertilizer synergists. Multiple studies have found that Harpins protein secreted by Gram-negative bacteria... [1] β-cryptogein secreted by Phytophthora cryptogaea [2] And the PeaT1 elicitor of *Alternaria tenuissima*. [3] Key signaling molecules can bind to receptors on the surface of plant cells, thereby activating the plant's own immune and growth systems, enhancing the plant's ability to resist pests, diseases, and adverse environments, and playing an important role in improving crop quality and increasing yield. [4] Among them, Harpins proteins have been studied the most extensively, and they are generally classified into four categories: Hpa1, HrpN, HrpW1, and HrpZ1. [1] It has been reported that exogenous application of Hpa1 and overexpression of the hrpZ gene can significantly induce an increase in the activity of plant defense-related enzymes and the expression of disease resistance-related genes, thereby improving the systemic resistance of the corresponding diseases. [5,6] Transgenic rice plants overexpressing the Harpin-encoding gene hrf1 exhibited enhanced drought tolerance via the ABA signaling pathway. [7] In addition, the study found that after soaking Arabidopsis seeds in HpaXpm protein of the Hpa1 family for 10 days, root length and plant fresh weight were significantly higher than those in the control group. [8] Rice seedlings treated with N-HrpE protein showed a significant increase in biomass compared to the control group, with root morphology parameters reaching their maximum values, demonstrating a clear growth-promoting effect. [9] .visible, Regardless of heterologous expression in plant cells Whether through the Harpins protein gene or exogenous spraying, Harpins protein can stimulate plants to produce a variety of beneficial effects, including increasing... Strengthening plant defense responses against pathogenic microorganisms, enhancing plant resistance to adverse conditions, and promoting plant growth and development, etc. Based on the above advantages, using Harpins protein as a fertilizer enhancer to produce green, high-efficiency fertilizers with more comprehensive nutrition, stronger targeting, and higher absorption and utilization rates has a promising application prospect.

[0004] Based on this, the present invention is proposed.

[0005] [References] [1]Choi MS, Kim W, Lee C, Oh C S. Harpins, multifunctional proteins secreted by gram-negative plant-pathogenic bacteria[J]. Molecular Plant-Microbe Interactions®, 2013, 26(10): 1115-1122. [2]Ricci P, Bonnet P, Huet JC, Sallantin M, Beauvais-Cante F, et al.Structure and activity of proteins from pathogenic fungi phytophthoraeliciting necrosis and acquired resistance in tobacco[J]. European Journal ofBiochemistry, 1989, 183(3): 555-563. [3] Zhang Wei. Study on PeaT1-induced disease resistance in tobacco system and its mechanism of action [D]. Chinese Academy of Agricultural Sciences, 2011: 24-40. [4] Liu Xueqiu, Zhang Huaqian, Du Chunmei. Protein-based plant immune inducers [J]. Chinese Journal of Biochemistry and Molecular Biology, 2024, 40(11): 1479-1492. [5]Wang D, Wang B, Wang J, Wang S, Wang W, et al. Exogenousapplication of harpin protein Hpa1 onto pinellia ternata induces systemicresistance against tobacco mosaic virus[J]. Phytopathology®, 2020, 110(6):1189-1198. [6]Du Q, Yang X, Zhang J, Zhong X, Kim K S, et al. Over-expression ofthe pseudomonas syringae harpin-encoding gene hrpZm confers enhancedtolerance to phytophthora root and stem rot in transgenic soybean[J].Transgenic Research, 2018, 27(3): 277-288. [7]Zhang L, Xiao S, Li W, Feng W, Li J, et al. Overexpression of aharpin-encoding gene hrf1 in rice enhances drought tolerance[J]. Journal ofExperimental Botany, 2011, 62(12): 4229-4238. [8]Liu Y, Zhou X, Liu W, Huang J, Liu Q, et al. HpaXpm, a novelharpin of xanthomonas phaseoli pv. manihotis, acts as an elicitor with highthermal stability, reduces disease, and promotes plant growth[J]. BMCMicrobiology, 2020, 20(1): 4. [9]Sheikh TMM, Haider MS, Hanif A, Ali H, Khan AR, et al. N-terminal HrpE from xanthomonas oryzae pv. oryzae mediates the regulation ofgrowth and photosynthesis in rice[J]. Plant Growth Regulation, 2022, 96(3):383-396. Summary of the Invention

[0006] This invention first relates to a signaling protein RH1, which is used to stimulate plant growth and increase crop yield. The plants include, but are not limited to, Arabidopsis thaliana and rapeseed. The polypeptide sequence of the signaling protein is shown in SEQ ID NO.1.

[0007] The present invention also relates to a nucleotide fragment encoding the signal protein RH1, preferably, the sequence of the nucleotide fragment is shown in SEQ ID NO.2.

[0008] The present invention also relates to compound fertilizers containing the signaling protein RH1.

[0009] Furthermore, the compound fertilizer contains: (1) One-thousandth (wt) of the signaling protein RH1; (2) Compound fertilizer substrate with N content of 15-30%, P2O5 content of 10-20%, and K2O content of 5-15%; In the compound fertilizer matrix, monoammonium phosphate serves as a phosphorus source to provide P2O5, urea serves as a nitrogen source to provide N, and potassium chloride serves as a potassium source to provide K2O. Preferably, the mass ratio of N, P2O5, and K2O in the compound fertilizer matrix is ​​15:6:9.

[0010] The present invention also relates to the application of the signaling protein RH1 or the compound fertilizer in the cultivation of crops, preferably, the crops being cabbage and rapeseed.

[0011] Furthermore, the present invention also relates to a method for cultivating rapeseed plants using the compound fertilizer, characterized in that 30-50 kg of the compound fertilizer is applied per mu of rapeseed, preferably 40 kg.

[0012] This invention also relates to the following applications of the aforementioned signaling protein RH1: (1) Upregulate the expression of phosphorus uptake genes in plants, and / or (2) Upregulate phosphorus transport genes in plants.

[0013] The present invention also relates to the use of the signaling protein RH1 in the preparation of formulations that upregulate phosphorus uptake genes and / or phosphorus transport genes in plants.

[0014] The present invention also relates to the application of the signaling protein RH1 in the preparation of fertilizer synergists, which are used to increase the crop growth promotion efficiency of existing fertilizers.

[0015] The beneficial effects of this invention are as follows: 1. In this study, a novel recombinant Harpins signaling protein, RH1, was constructed by extracting functional fragments of Harpins proteins from various pathogens and recombining them.

[0016] 2. The signaling protein RH1 was prepared into a compound fertilizer, and its biological functions were investigated in Arabidopsis thaliana and Brassica napus L., respectively. This signaling protein RH1 can serve as a low-cost, highly efficient, and safe fertilizer synergist to improve fertilizer utilization, increase crop yield and quality, and provide theoretical basis and technical support for the continued expansion of the application of bioactive substances in agriculture in my country, thus promoting the achievement of green agricultural development goals.

[0017] 3. Previous studies on harpins proteins have mostly focused on enhancing crop disease resistance through transgenic or external application, with few reports on their use as fertilizer synergists. This study applied signal protein RH1 compound fertilizer to rapeseed in a field trial. The results showed that the rapeseed treated with signal protein RH1 compound fertilizer exhibited better growth than those treated with ordinary compound fertilizer, and the yield per mu (a Chinese unit of area, approximately 0.067 hectares) was increased compared to the control group. Among the three treatment groups, the highest yield increase rate reached 58.05%, indicating that signal protein RH1 compound fertilizer has a significant yield-increasing effect. Regarding the relationship between fertilizer application rate and rapeseed yield, the highest yield was observed with 40 kg / mu of signal protein RH1 compound fertilizer (234.88 kg / mu). Increasing the fertilizer application rate to 50 kg / mu resulted in a decrease in yield (197.35 kg / mu), suggesting that 40 kg / mu of signal protein RH1 compound fertilizer is more suitable. The three control groups treated with ordinary compound fertilizer did not show a peak yield, indicating that their optimal fertilizer application rate may be greater than 50 kg / mu. Therefore, signal protein RH1 compound fertilizer has a significant effect of reducing fertilizer use while increasing yield. Attached Figure Description

[0018] Figure 1 The signaling protein RH1 promotes the growth and development of Arabidopsis thaliana. 1A: Arabidopsis thaliana treated with 0.5 g / L ordinary compound fertilizer (left) and signal protein RH1 compound fertilizer (right) for 10 days; 1B-D: Total biomass, aboveground biomass, and belowground biomass of Arabidopsis thaliana after 10 days of treatment; 1E-G: Root phenotype (scale bar = 1 cm), root length and inorganic phosphorus content of Arabidopsis thaliana after 10 days of treatment; 1H-K: Arabidopsis thaliana was treated with 0.5 g / L of ordinary compound fertilizer culture medium and compound fertilizer culture medium containing signal protein RH1 for 6 h, and the expression levels of genes related to phosphorus absorption and transport were detected.

[0019] Figure 2 Signal protein RH1 promotes the growth and development of Brassica napus. 2A: Brassica napus was treated with 0.5 g / L ordinary compound fertilizer (left) and compound fertilizer containing signal protein RH1 (right) for 10 days (top) and 20 days (bottom); 2B: Root phenotype of rapeseed 10 days after treatment (scale bar = 1 cm); 2C-G: Total biomass, aboveground biomass, underground biomass, root length, and inorganic phosphorus content of rapeseed after 20 days of treatment; 2H-O: After treating rapeseed with 0.5 g / L ordinary compound fertilizer culture medium and compound fertilizer culture medium containing signal protein RH1 for 6 h, the expression levels of genes related to phosphorus absorption and transport were detected.

[0020] Figure 3 Field growth of "Kaiyu 118" under different fertilization treatments (T1-T6). Detailed Implementation

[0021] 1. Construction of the signaling protein RH1 The recombinant protein fragments were derived from Harpins proteins secreted by *Ralstonia solanacearum*, *Erwinia amylovora*, *Erwinia pyrifoliae*, *Pseudomonas syringae pv. Syringae*, and *Ralstonia pseudosolanacearum*, with GenBank accession numbers EF080949.1, U94513.1, WP_012669299.1, L14775.1, and WP_221743891.1, respectively.

[0022] The amino acid sequence of the recombinant protein is shown in SEQ ID NO.1, and the sequence encoding the nucleotide fragment is shown in SEQ ID NO.2. SEQ ID NO.1 (amino acid sequence): TSSSPGLFQSGGDNGLGGHNANSALGQQPIDRQTIEQMAQLLAELLKSLLDGTGIGMKAGIQALNDIGTHSDSSTRSFVNKGDRAMAKEIGQFMDQYPEVFGKPQYQKGPGQEVKTDDKSWAKALSKPDDDGMTPASMEQFNKAKGMIKSAMAGDTGNGNLQARGAGGSSLGIDAMMAGDAINNMALGKLGAADGEKLGDNFGASADSASGTGQQDLMTQVLNGLAKSMLDDLLTKQDGGTSFSEDDMPMLNKIAQDGMSIQIDRPNNHFQTPSTWNHDAGSSIDTSQLQRAVQLLDQVLQQLEARKLFGNMLNQPGADNAGQNHAGGHGGGHHGGSNGFGENGRFGSDGGGLGTPVNTPQTGTSANGGQSAQDLDQLLGGLLLKGLEATLKD SEQ ID NO.2 (Nucleic acid sequence): 2. Recombinant expression and purification of signal protein RH1 Using conventional molecular biology methods in this field, with Escherichia coli BL21 as the host bacterium and plasmid pET-28a as the expression vector, a recombinant expression strain of the signal protein RH1 was constructed through gene cloning, transformation, and screening verification.

[0023] 2.1 Protein Expression Activation of RH1 signaling protein expression strain: Inoculate the strain at 1% into a shake flask containing 50 mL LB medium, add kanamycin to a final concentration of 25 μg / mL, and incubate overnight at 37°C and 200 rpm.

[0024] The next day, 1% was inoculated into a shake flask containing 1L of TB medium, and kanamycin was added to a final concentration of 25ug / mL. The flask was incubated at 37℃ and 200 rpm for 3-4 hours. The temperature was then lowered to 20℃, and IPTG was added to a final concentration of 0.5 mM. The flask was then incubated for another 16 hours.

[0025] Collect the bacterial cells by centrifuging the cultured bacterial solution at 5000 rpm for 10 min at 4 °C. Discard the supernatant and use the obtained bacterial cells immediately, or store them at -80 °C for long-term storage.

[0026] 2.2 Ni-Sepharose HP affinity purification of proteins The collected bacterial cells were resuspended in 1×NPI 10 Buffer, and PMSF was added to a final concentration of 1 mM to inhibit protease activity. After sonication, the cells were centrifuged at 20,000 rpm for 30 min at 4 ℃. The supernatant was collected, filtered through a 0.22 μm filter membrane, and loaded onto a Ni-NTA chromatography column (Ni-Sepharose HP, GE Healthcare, packed in an XK 16 / 20 column, 20 mL column volume).

[0027] After loading the sample, continue washing the chromatography column with 1×NPI 10 Buffer for 5 column volumes. After the UV absorbance value decreases to a stable level, perform stepwise gradient elution using NPI-250 Buffer.

[0028] Samples of the supernatant, flow-through, and eluted protein solution were taken and analyzed by SDS-PAGE.

[0029] 3. Test materials and location Plant materials: Arabidopsis thaliana WT (Columbia), Brassica napus WT (Westar), and rapeseed varieties “Kaiyu 118” and “Huayouza 62”.

[0030] Fertilizer: Ordinary compound fertilizer (N-P2O5-K2O: 15-6-9) provided by Hubei Weishengyuan Biotechnology Co., Ltd. This 15-6-9 compound fertilizer is a low-concentration chlorine-based compound fertilizer. The company uses monoammonium phosphate as the phosphorus source, combined with domestically produced urea (nitrogen) and potassium chloride (potassium), and adds humic acid / organic matter as a synergist. Granulation is then performed using the agglomeration method. A simple formula example (1 ton): 326 kg of urea (46% N, providing approximately 15 kg of N) 136 kg of monoammonium phosphate (44% P2O5, providing approximately 6 kg of P2O5) 150 kg of potassium chloride (60% K2O, providing approximately 9 kg of K2O) 80 kg of dry humic acid (to improve utilization and blacken granules) Fill the filler (dolomite / bentonite) to 1 t (approximately 308 kg - also for adjusting acidity and alkali and preventing caking).

[0031] The production process of compound fertilizer can be carried out according to conventional methods in this field, and the general process route is as follows: Raw material crushing → metering and mixing → rotary drum granulation (steam injection + acid) → drying → sieving → cooling → coating → packaging into 25 kg small packages.

[0032] In the production process of compound fertilizer containing signal protein RH1, simply add 1 / 1000 of signal protein RH1 (protein powder) during the above coating process. The two compound fertilizers are identical in all components except for the signaling protein.

[0033] Address: Wutonghu New District, Ezhou City, Hubei Province (114°58´E, 30°40´N), Danjiangkou City, Shiyan City, Hubei Province (111°50´E, 32°54´N).

[0034] 4. Test treatment and index determination Arabidopsis thaliana WT: After sterilization, Arabidopsis thaliana seeds were laid on MS medium for germination. After 6 days, they were transferred to vermiculite and irrigated with 0.5 g / L ordinary compound fertilizer and signal protein RH1 compound fertilizer, respectively. After 10 days of treatment, phenotype and root length were recorded. Biomass (based on fresh weight) was measured in groups of 10 Arabidopsis thaliana plants. The inorganic phosphorus content was determined by the phosphomolybdic blue colorimetric method using 0.1 g of fresh sample. Plant culture was carried out at room temperature (25-30℃) under 16 h light / 8 h dark conditions.

[0035] Brassica napus (WT) seeds were directly sown in vermiculite. After germination, the seeds were irrigated with either 0.5 g / L ordinary compound fertilizer or RH1 signal protein compound fertilizer. Phenotypic results and root length were recorded after 10 days of treatment. Individual plant biomass (based on fresh weight) was measured after 20 days. Inorganic phosphorus content was determined using the phosphomolybdic blue colorimetric method with 0.1 g of fresh sample. Plant cultures were conducted at room temperature (25-30℃) under 16 h light / 8 h dark conditions.

[0036] 5. Gene expression detection Six-day-old Arabidopsis thaliana rootlets (WT) and Brassica napus rootlets (WT) were transferred to 0.5 g / L ordinary compound fertilizer culture medium and signal protein RH1 compound fertilizer culture medium, respectively, and treated for 6 h. Total RNA was extracted using the Trizol method, followed by reverse transcription to obtain cDNA. The expression of relevant phosphorus uptake and transport genes was detected by RT-qPCR. The gene amplification primers are shown in Table 1.

[0037] Table 1. Primer sequence information for RT-qPCR amplification

[0038] 6. Field trials Wutonghu New District, Ezhou City, Hubei Province: The experiment set up two treatments: ordinary compound fertilizer and signal protein RH1 compound fertilizer, and three compound fertilizer application rates of 25 kg / mu, 40 kg / mu, and 50 kg / mu, for a total of 6 fertilization treatments (T1~T6, Table 2).

[0039] Table 2. Compound fertilizer formula and application rate for the "Kaiyu 118" experimental treatment groups

[0040] The rapeseed field was divided into 6 experimental zones, each with an area of ​​25m². 2 (2.5m × 10m), totaling 225 m 2 After evenly spreading the fertilizer for each treatment onto the ground, sow "Kaiyu 118" rapeseed seeds and thin the seedlings when they have 3 leaves and 1 heart.

[0041] Except for fertilizer treatment, all other agricultural operations are the same. Yield is measured when the rapeseed matures, using a three-point sampling method to select three points in each experimental area, with each point containing a 1-meter sample. 2 The harvested rapeseed was individually packaged, dried thoroughly, threshed, and then weighed.

[0042] In Danjiangkou City, Shiyan City, Hubei Province, two treatments were set up: 40 kg / mu of ordinary compound fertilizer (T3 treatment group) and 40 kg / mu of RH1 enhanced compound fertilizer (T4 treatment group). The rapeseed field was divided into two experimental areas, each with an area of ​​2 mu (667 m²). 2 / mu), totaling 4 mu.

[0043] After evenly spreading the two fertilizer treatments on the field, "Huayouza 62" rapeseed seeds were sown. Except for the fertilizer treatments, all other agricultural operations were the same. Yield was measured at rapeseed maturity using a five-point sampling method, selecting five points in each experimental plot, with each point containing a 1 m sample. 2 The harvested rapeseed was individually packaged, dried thoroughly, threshed, and then weighed.

[0044] 7. Data Processing The statistical data was initially organized using Excel 2021, and then significance tests and analysis of variance were performed using SPSS 17.0. Graphs were created using Adobe Photoshop 2022 and GraphPad Prism 9. Example 1: Effects of signaling protein RH1 on plant growth and development 1. Effects of signaling protein RH1 on the growth and development of Arabidopsis thaliana Six-day-old Arabidopsis thaliana WT was transplanted into vermiculite and irrigated with ordinary compound fertilizer and compound fertilizer containing signal protein RH1, respectively.

[0045] The results show that Arabidopsis thaliana irrigated with RH1 signal protein compound fertilizer (3g of compound fertilizer added to a 500ml pot) showed significantly better growth than the control group. Their total biomass, aboveground biomass, underground biomass, and root length increased by 80.94%, 80.65%, 82.95%, and 79.80%, respectively, compared to the control group. Figure 1 AF). The results of inorganic phosphorus content determination showed that the inorganic phosphorus content in both the aboveground and underground parts of Arabidopsis thaliana irrigated with RH1 signal protein compound fertilizer was higher than that in the control group (AF). Figure 1 G), and its corresponding Phosphorus absorption Gene PHT1;1、PHT1;4 Phosphorus transport genes PHO1 Expression levels were upregulated, among which PHT1;4 The most significant increase was in the upward adjustment (approximately 14 times). Figure 1 HK).

[0046] The results indicate that the signaling protein RH1 can promote the absorption and transport of inorganic phosphorus in Arabidopsis thaliana, thereby promoting its growth and development.

[0047] 2. Effects of signaling protein RH1 on the growth and development of Brassica napus The results of treating rapeseed seedlings with compound fertilizer containing the signal protein RH1 were similar to those of Arabidopsis thaliana.

[0048] Signal protein RH1 compound fertilizer (10g compound fertilizer added to a 500ml pot after potting) significantly promoted the increase of rapeseed seedling biomass and root development. The total biomass, aboveground biomass, underground biomass, and root length increased by 76.57%, 74.16%, 95.78%, and 39.15% respectively compared to the control group. Figure 2 AF). The results of inorganic phosphorus content determination showed that the inorganic phosphorus content in both the aboveground and underground parts of rapeseed seedlings treated with compound fertilizer containing signal protein RH1 was higher than that in the control group. Figure 2 G), and the RH1 signal protein compound fertilizer treatment group Phosphorus absorption gene BnA09a.PHT1;1, BnA09b.PHT1;1, BnC04.PHT1;4, BnC08.PHT1;4 and Phosphorus transporter genes BnA01.PHO1, BnC01.PHO1, BnC06.PHO1;H1, BnA07.PHO1;H1 Expression levels were significantly upregulated ( Figure 2 HO).

[0049] The above results indicate that the signaling protein RH1 can also promote the absorption and transport of inorganic phosphorus in rapeseed, thereby promoting its growth and development.

[0050] Example 2: Effects of different fertilizers and application rates on rapeseed yield The use of fertilizer enhancers is one of the key ways to reduce fertilizer use and increase efficiency in agricultural production, and it is of great significance to promoting the sustainable development of agricultural production.

[0051] To investigate the role of the signaling protein RH1 in actual production, this study conducted a field experiment on rapeseed variety "Kaiyu 118" in Wutonghu New District, Ezhou City, Hubei Province. The experiment was divided into 6 fertilization treatments (Table 2). Except for the different fertilizers added, the other agricultural operations were the same.

[0052] See results Figure 3 As can be seen from Table 3, The rapeseed growth of the treatment groups (T2, T4, T6) treated with compound fertilizer containing signal protein RH1 was better than that of the control group (T1, T3, T5).

[0053] The yield measurement results show that The rapeseed yield per mu (a Chinese unit of area, approximately 0.067 hectares) increased in all treatment groups treated with signal protein RH1 compound fertilizer compared to the control group. The T4 treatment group (40 kg / mu of signal protein RH1 compound fertilizer) had the highest rapeseed yield, averaging 234.88 kg / mu, followed by the T6 treatment group (50 kg / mu of signal protein RH1 compound fertilizer), with an average yield of 197.35 kg / mu (Table 3). Significant differences in rapeseed yield were observed between treatment groups T4 and T1, T2, T3, and T5, while no significant differences were found in the yields of the other treatment groups.

[0054] Considering the production requirements of reducing fertilizer use and increasing efficiency, the optimal fertilization treatment is 40 kg / mu of compound fertilizer containing signal protein RH1. .

[0055] The same T4 fertilization scheme was also used in a large-scale field trial of the rapeseed variety "Huayouza 62" to achieve ideal yield increase (average yield of 200.51 kg per mu), which is 28.66% higher than the T3 fertilization scheme (Table 4).

[0056] Table 3 Yield of "Kayu 118" under different fertilization treatments

[0057] Note: Different lowercase letters (a\b\ab) indicate the significance of the difference between groups at the P < 0.05 level (no significant difference between groups with the same lowercase letter, and significant difference between groups with different letters).

[0058] Table 4 Yield of "Huayouza 62" under different fertilization treatments

[0059] Note: Different lowercase letters indicate significant differences between treatments at the P < 0.05 level.

[0060] Finally, it should be noted that the above embodiments are only used to help those skilled in the art understand the technical content of the present invention, and are not intended to limit the scope of protection of the present invention.

Claims

1. A signal protein RH1, the polypeptide sequence of which is shown as SEQ ID NO. 1; the signal protein RH1 is used for stimulating plant growth and increasing crop yield, the plants including but not limited to Arabidopsis thaliana and Brassica napus.

2. A nucleotide fragment encoding the signal protein RH1 of claim 1, preferably the sequence of the nucleotide fragment is shown as SEQ ID NO.

2.

3. A compound fertilizer containing the signal protein RH1 of claim 1.

4. The compound fertilizer according to claim 3, characterized in that, The compound fertilizer comprises: (1) one thousandth (wt) of the signal protein RH1; (2) a compound fertilizer base with N content of 15-30%, P2O5 content of 10-20%, and K2O content of 5-15%.

5. The compound fertilizer according to claim 4, characterized in that, In the compound fertilizer base, monoammonium phosphate provides P2O5 as a phosphorus source, urea provides N as a nitrogen source, and potassium chloride provides K2O as a potassium source. Preferably, in the compound fertilizer base, the mass ratio of N, P2O5, and K2O is 15:6:

9.

6. The signal protein RH1 of claim 1, the nucleotide fragment of claim 2, or the compound fertilizer of claims 3-5 for use in cultivating crops, preferably the crops are Brassica oleracea and Brassica napus.

7. A method of growing Brassica plants using the compound fertilizer of claim 5, characterized in that, 30-50 kg of the compound fertilizer is applied per mu of Brassica napus.

8. The signal protein RH1 of claim 1 for use in: (1) up-regulating the expression of a phosphorus uptake gene in a plant, and / or (2) up-regulating a phosphorus transport gene in a plant.

9. The signal protein RH1 of claim 1 for use in the preparation of a preparation for up-regulating a phosphorus uptake gene and / or a phosphorus transport gene in a plant.

10. The signal protein RH1 of claim 1 for use in the preparation of a fertilizer synergist for increasing the crop growth promoting efficiency of an existing fertilizer.

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