Paecilomyces variotii in improving salt-alkali stress resistance of wheat and a wheat planting method in saline-alkali land

Abstract

This invention discloses the application of *Penicillium wani* in improving wheat's resistance to salt-alkali stress and a method for planting wheat in saline-alkali land, belonging to the field of crop cultivation technology. This invention has the advantages of simple operation, rapid processing, and excellent results. The specific method is as follows: during the sowing period, wheat seeds are treated with a low concentration (1 μg / ml) of *Penicillium wani* solution. This treatment can improve wheat's resistance to salt stress at concentrations up to 0.3% and salt-alkali stress at 75 mM (pH 9.2), thereby increasing wheat yield. This invention is applicable to the cultivation of wheat and other crops and has broad prospects for widespread application.

Description

Technical Field

[0001] This invention belongs to the field of crop cultivation technology, specifically relating to the application of Wan's penicillin in improving wheat's resistance to salt and alkali stress and a method for planting wheat in saline-alkali land. Background Technology

[0002] The statements herein provide only background information in relation to this invention and do not necessarily constitute prior art.

[0003] Soil salinization refers to the natural geological process by which salts accumulate on the soil surface, forming saline soil. It mainly occurs in arid, semi-arid, and semi-humid climate zones, as well as coastal lowlands affected by seawater intrusion. Soil salinization leads to soil compaction and decreased fertility, hindering crop nutrient absorption and thus impeding crop growth, severely impacting agricultural production.

[0004] Unlike salt stress, which originates from neutral salts such as sodium chloride and sodium sulfate and causes difficulties in water absorption by plant roots through osmotic stress, as well as the entry of sodium and chloride ions into the plant body, interfering with metabolism, salt-alkali stress originates from alkaline salts such as sodium bicarbonate and sodium carbonate. In addition to the two types of damage caused by salt stress, it also has another core destructive factor—high pH. This factor directly corrodes the root system, damaging the structure and function of root cells. Simultaneously, a high pH environment reduces the solubility of many mineral nutrients in the soil (such as iron and phosphorus), causing an imbalance in plant mineral nutrition and exacerbating the damage. Existing research confirms that plants employ different strategies to cope with salt stress and salt-alkali stress, with salt-alkali stress generally causing greater harm.

[0005] In saline-alkali stress environments, seed germination and seedling growth are relatively crucial stages. Therefore, exploring the response mechanisms of seed germination under saline-alkali stress is of great significance for the development and utilization of saline-alkali soils. In recent years, there has been a large amount of research on the response mechanisms of salt stress to crop seed germination and seedling growth, while research on saline-alkali stress itself is relatively insufficient. Currently, the focus on addressing the problem of saline-alkali land is on cultivating salt-tolerant crops, but this approach faces challenges such as long research cycles, high costs, and significant difficulties.

[0006] Wheat is an important food crop. However, salt-tolerant wheat germplasm resources are scarce in China, and scientific and technological breakthroughs are urgently needed.

[0007] Saline-alkali soil improvement technologies mainly include a diversified system based on physical, chemical, and biological methods. Physical improvement technologies directly alter soil structure and water-salt dynamics through mechanical means. Deep plowing and tilling, as traditional measures, can break up the salt accumulation layer and enhance permeability. Physical methods, as a fundamental improvement approach, primarily adjust soil structure and water-salt dynamics through mechanical operations. However, their effectiveness is limited by the complexity of the initial terrain and subsequent management measures. Chemical improvement methods focus on adjusting soil ionic composition and pH balance by adding specific substances to quickly alleviate saline-alkali damage. Gypsum (CaSO4·2H2O), as a traditional amendment, uses Ca... 2+ Replace Na in soil colloids + It promotes sodium salt leaching and improves soil structure. Applying desulfurized gypsum increases the content of large aggregates and porosity in saline-alkali soils, significantly improving water infiltration efficiency. Humic acids, due to their colloidal properties, can complex salt ions and enhance soil water-holding capacity. Aluminum sulfate, through hydrolysis, produces Al... 3+ Replacement of Na + It also forms hydroxyaluminum polymers in the soil, promoting the formation of aggregates. However, chemical amendments may introduce pollutants such as heavy metals.

[0008] A prior patent (CN109824757A) first disclosed the structure of the cyclic (isoleucine-isoleucine) dipeptide. Recently, this bio-derived plant growth regulator was granted the Chinese generic name "Wanshi Penicillin". This compound has the effect of improving the germination rate of wheat seeds under salt stress, promoting root length and number, and has significant effects in inducing flowering, resisting cucumber downy mildew, resisting frost, increasing yield, and improving fertilizer utilization. However, whether it has a beneficial effect on the growth of wheat seedlings under salt-alkali stress is still unknown.

[0009] Effective methods are still needed to improve wheat growth under salt and alkali stress. Summary of the Invention

[0010] In view of the shortcomings of the existing technology, the purpose of this invention is to provide the application of Wan's penicillin in improving the resistance of wheat to salt and alkali stress and a method for planting wheat in saline-alkali land.

[0011] The above-mentioned objectives are achieved by the present invention through the following technical solution: In a first aspect, the present invention provides the application of Wan's penicillin in improving the resistance of wheat to salt and alkali stress.

[0012] The comparative study of this invention found that adding penicillin in the form of seed dressing during wheat vernalization has a significant effect in alleviating salt-alkali stress and can significantly improve wheat survival rate.

[0013] The comparative study of this invention found that adding penicillin to the roots of wheat seedlings can improve the resistance of wheat to salt and alkali stress and significantly improve the core indicators of wheat under salt and alkali stress, such as plant height, chlorophyll content, and yield.

[0014] Furthermore, the concentration of Wan's penicillin added is not less than 1 μg / ml.

[0015] Furthermore, the salt-base stress condition is a sodium carbonate-sodium bicarbonate buffer solution.

[0016] Furthermore, in the aforementioned salt-alkali stress conditions, the pH value of the 75 mM sodium carbonate-sodium bicarbonate buffer solution is 9.0-10.0.

[0017] The purpose of this invention is to significantly improve the resistance of cereal plants to salt and alkali stress and significantly increase wheat yield by treating seeds with a low concentration (1 μg / ml) of penicillin or by drenching seedlings with penicillin.

[0018] Furthermore, this invention is the first to discover that treatment with *Penicillium wani* can improve the survival rate of wheat under salt-alkali stress. Compared with the control (H2O), the survival rate of wheat treated with *Penicillium wani* solution under salt-alkali stress was significantly improved, indicating that treatment with *Penicillium wani* can enhance the resistance of wheat to salt-alkali stress. This substance can be applied to the cultivation of wheat crops.

[0019] In the embodiments section of this invention, the applicant describes a method for improving the salt and alkali stress resistance of wheat crops using Wan's penicillin.

[0020] Furthermore, the wheat is selected from jointed wheat and common wheat.

[0021] Furthermore, the common wheat is selected from at least one of Jimai 22, B6-3-3 and Yannong 999.

[0022] Comparative experiments revealed that different wheat varieties responded differently to penicillin.

[0023] Among them, applying 1 μg / ml of penicillin to B6-3-3 and Yannong 999 (YN999) can completely reverse salt-alkali stress and promote the growth of B6-3-3 and Yannong 999 under salt-alkali stress conditions, thereby increasing wheat yield, including significantly increasing plant height, chlorophyll content, thousand-grain weight, grain length, and grain width under salt-alkali stress.

[0024] Furthermore, the application of 1 μg / ml of penicillin can completely restore the growth of wheat under saline-alkali conditions.

[0025] Furthermore, the application of penicillin wannabese under saline-alkali stress conditions can increase the yield of wheat, wherein the wheat is selected from Jimai 22, B6-3-3 and Yannong 999.

[0026] Secondly, the present invention provides a method for planting wheat in saline-alkali land, comprising the following steps: S1, wheat seeds germinate at room temperature; S2, Low-temperature continuous light treatment of wheat seedlings: After germination at room temperature for 2-5 days, wheat seedlings are moved to a low-temperature environment of 2-7℃ for 24-hour light treatment and vernalization treatment for 30-35 days. S3, transplant the vernalized wheat seedlings into the soil and treat the roots with Wan's penicillin solution.

[0027] Preferably, in step S3, a solution of penicillin with a concentration of 1 μg / ml is used for root irrigation.

[0028] Preferably, the wheat is jointed wheat, Jimai 22, or B6-3-3.

[0029] The beneficial effects achieved by one or more embodiments of the present invention described above are as follows: Wan's penicillin, also known as cyclic (isoleucine-isoleucine) dipeptide, significantly enhances the resistance of wheat and its close relative, *Gnaphalium affine*, to salt-alkali stress, thereby increasing wheat yield in saline-alkali environments. Treating wheat seeds with Wan's penicillin through seed dressing or root irrigation can improve wheat resistance to salt-alkali stress, providing a potential approach for increasing wheat yield in saline-alkali lands. Importantly, the resistance mediated by Wan's penicillin is broad-spectrum across different wheat varieties and their close relatives, suggesting a wide range of potential applications for Wan's penicillin. Attached Figure Description

[0030] The accompanying drawings, which form part of this invention, are used to provide a further understanding of the invention. The illustrative embodiments of the invention and their descriptions are used to explain the invention and do not constitute an improper limitation of the invention.

[0031] Figure 1 The purpose of this study is to enhance the resistance of *Gnaphalium affine* to salt and alkali stress using *Penicillium wansi* in Example 1.

[0032] Figure 2 In Example 2, the seed dressing with *Penicillium wansii* was used to improve the resistance of common wheat to salt and alkali stress.

[0033] AB: In the hydroponic experiment, seed treatment with 1 μg / ml of *Penicillium wilfordii* improved the survival of wheat variety Jimai 22 under salt-alkali stress (75 mM); CD: In the hydroponic experiment, seed treatment with 1 μg / ml of *Penicillium wilfordii* improved the survival of wheat variety B6-3-3 under salt-alkali stress (75 mM); E: Survival rate of common wheat seedlings in AD.

[0034] Figure 3The root irrigation treatment with penicillin wandii in Example 3 was used to increase the plant height of ordinary wheat.

[0035] AE: Common wheat varieties Jimai 22 (A), B6-3-3 (B), Yannong 999 (C), Shannong 116 (D) and Fielder (E) were transplanted into the soil after 30 days of vernalization. One week after transplanting, they were treated with water (CK), salt stress (75mM) + different concentrations (0, 1, 10, 100, 1000 ng / ml) of Wanshi penicillin, respectively, for root irrigation.

[0036] Figure 4 The root irrigation treatment of common wheat seedlings with *Penicillium wani* in Example 4 was used to improve the resistance of common wheat to salt and alkali stress.

[0037] AD: Common wheat varieties Jimai 22 (A), B6-3-3 (B), Yannong 999 (C) and Zhongguochun (D) were transplanted into the soil after 30 days of vernalization. One week after transplanting, they were treated with water (CK), salt-alkali stress (75mM), salt-alkali stress (75mM) + Wanshi penicillin (1μg / ml), and Wanshi penicillin (1μg / ml) respectively for root irrigation.

[0038] Figure 5 The root irrigation treatment with Wan's penicillin in Example 4 was used to increase the yield of ordinary wheat.

[0039] AC: Common wheat varieties Jimai 22 (A), B6-3-3 (B) and Yannong 999 (C) were transplanted into the soil after 30 days of vernalization. One week after transplanting, they were treated with water (CK), salt stress (75mM), salt stress (75mM) + Wan's penicillin (1μg / ml) and Wan's penicillin (1μg / ml) respectively for root irrigation. Detailed Implementation

[0040] It should be noted that the following detailed description is illustrative and intended to provide further explanation of the invention. Unless otherwise specified, all technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0041] Measurement indicators and measurement methods 1. Jointed goatgrass growth index: Five replicates were taken for each treatment, and the number of jointed goatgrass tillers was counted and recorded as the jointed goatgrass growth index.

[0042] 2. Survival rate: 10 replicates were taken for each treatment. The number of green surviving common wheat and the number of withered and necrotic common wheat were counted. The percentage of green surviving common wheat out of the total number was calculated and recorded as the survival rate of common wheat.

[0043] 3. Chlorophyll content determination: Cut fresh leaves into small pieces. Accurately weigh 0.1-0.2 g (recorded as W, unit: g) of sample and place it in a mortar. Add 2-3 mL of extraction solvent to the mortar. Grind thoroughly until the chlorophyll is completely dissolved. Filter the extract and dilute to 10 mL in a volumetric flask. Pour an appropriate amount of chlorophyll extract into a cuvette and measure its absorbance at wavelengths of 663 nm (A663) and 645 nm (A645), respectively.

[0044] Total chlorophyll concentration (mg / L): Ctotal = Ca + Cb = 20.29 × A645 + 8.02 × A663.

[0045]

[0046] 4. Plant height measurement: 30 days after transplanting, 10 ordinary wheat plants were randomly selected, and the height from the base of the wheat plant to the top of the ear was measured with a ruler. The average value was calculated.

[0047] 5. Determination of wheat grain length (GL): Using dried grains as the standard, five replicates were taken for each treatment. Ten grains were randomly selected from each replicate and arranged end to end. The total length was measured with a ruler, and the average length of the grains was calculated and recorded as the wheat grain length.

[0048] 6. Determination of wheat grain width (GW): Using dried grains as the standard, five replicates were taken for each treatment. Ten grains were randomly selected and arranged side by side. The total width was measured with a ruler, and then the average width of the grains was calculated and recorded as the wheat grain width.

[0049] 7. Determination of thousand-grain weight (TGW): Using dried grains as the standard, three replicates were taken for each treatment, and 50 grains were randomly selected from each replicate for weighing. The average value was calculated and then converted to the thousand-grain weight, which was recorded as the thousand-grain weight of ordinary wheat grains.

[0050] The present invention will be further described below with reference to the embodiments.

[0051] Example 1: Wan's penicillin enhances the resistance of jointed goatgrass to salt-alkali stress (1) Overview of the test site and test materials The test site is located at Building B5, No. 3366 Longteng Road, Beijipo Street, Daiyue District, Tai'an City, Shandong Province.

[0052] Jointed barley: JJM-4.

[0053] Salt-alkali stress treatment: Salt-alkali stress was treated by drenching the roots with 50 ml of 75 mM sodium carbonate-sodium bicarbonate buffer (pH 9.0-10.0). Sodium carbonate and sodium bicarbonate were purchased from Sigma-Aladin (USA).

[0054] (2) Experimental Design Seeds of the jointed goatgrass variety were placed in a petri dish lined with filter paper, and an appropriate amount of water was added. The seeds were then allowed to germinate at room temperature.

[0055] Jointed wheat seedlings subjected to 30 days of continuous low-temperature light treatment: After germination at room temperature for 2 days, the wheat seedlings were moved to a 4℃ low-temperature environment for 24-hour light treatment and vernalization treatment for 30 days.

[0056] Vernalized jointed goatgrass seedlings were transplanted into plastic pots (9×9×10 cm) filled with nutrient soil, with 5 seedlings transplanted into each pot. Seven days later, the roots were drenched with a 75 mM sodium carbonate-sodium bicarbonate buffer solution (pH 9.0-10.0), with different concentrations (0, 10, 100, 1000 ng / ml) of penicillin added. The growth of the jointed goatgrass was recorded after 14 days of treatment.

[0057] The results are as follows Figure 1 As shown, under salt-alkali stress provided by 75 mM sodium carbonate-sodium bicarbonate buffer (pH 9.0-10.0), the growth indicators of jointed goatgrass decreased significantly. Root drenching with different concentrations (10, 100, 1000 ng / ml) of penicillin significantly improved the growth of jointed goatgrass under salt-alkali stress. In particular, root drenching with 1000 ng / ml penicillin completely reversed the inhibitory effect of salt-alkali stress on jointed goatgrass.

[0058] Example 2, Hydroponic Experiment: Wan's penicillin enhances the resistance of common wheat to salt-alkali stress. (1) Overview of the test site and test materials The test site is located at Building B5, No. 3366 Longteng Road, Beijipo Street, Daiyue District, Tai'an City, Shandong Province.

[0059] Common wheat: Jimai 22 (JM22), B6-3-3.

[0060] Salt-alkali stress treatment: wheat germinating seedlings were treated with 20 ml of 75 mM sodium carbonate-sodium bicarbonate buffer (pH 9.0-10.0). Sodium carbonate and sodium bicarbonate were purchased from Sigma-Aladin (USA).

[0061] (2) Experimental Design Common wheat germination: Common wheat seeds are placed in a petri dish lined with filter paper, an appropriate amount of water is added, and they are germinated at room temperature.

[0062] Salt-alkali stress treatment during vernalization: Treatment 1: Two days after the wheat germinated, 20 ml of 75 mM sodium carbonate-sodium bicarbonate buffer (pH 9.0-10.0) was added to a petri dish and placed in a 4°C refrigerator for vernalization treatment for 30 days.

[0063] Treatment 2: Two days after the wheat germinated, 20 ml of 75 mM sodium carbonate-sodium bicarbonate buffer (pH 9.0-10.0) was added to a petri dish, along with 1 μg / ml of penicillin, and the dish was placed in a 4°C refrigerator for 30 days of vernalization.

[0064] Common wheat seedlings that had undergone vernalization were transplanted into plastic flowerpots (9×9×10 cm) filled with nutrient soil, with 5 seedlings transplanted into each flowerpot. The growth of the common wheat was recorded 14 days after transplanting.

[0065] The results are as follows Figure 2 As shown in Figures A to E, under saline-alkali stress, the growth of Jimai 22 was completely inhibited. Seed treatment with 1 μg / ml of penicillin significantly improved the survival rate of Jimai 22 and B6-3-3, with the survival rate of B6-3-3 reaching about 80%. Example 3, Soil cultivation experiment: Wan's penicillin improves the resistance of common wheat to salt and alkali stress.

[0066] (1) Overview of the test site and test materials The test site is located at Building B5, No. 3366 Longteng Road, Beijipo Street, Daiyue District, Tai'an City, Shandong Province.

[0067] Common wheat varieties: Jimai 22, B6-3-3, Yannong 999, Shannong 116, Fielder.

[0068] Salt-alkali stress treatment: Salt-alkali stress was achieved by drenching the roots of vernalized wheat seedlings with 50 ml of 75 mM sodium carbonate-sodium bicarbonate buffer (pH 9.0-10.0). Sodium carbonate and sodium bicarbonate were purchased from Sigma-Aladin (USA).

[0069] (2) Experimental Design Common wheat germination: Common wheat seeds are placed in a petri dish lined with filter paper, an appropriate amount of water is added, and they are germinated at room temperature.

[0070] Jointed wheat seedlings subjected to 30 days of continuous low-temperature light treatment: After germination at room temperature for 2 days, the wheat seedlings were moved to a 4℃ low-temperature environment for 24-hour light treatment and vernalization treatment for 30 days.

[0071] Salt-alkali stress after transplanting: Vernalized common wheat seedlings were transplanted into plastic pots (9×9×10 cm) filled with nutrient soil, with 5 seedlings per pot. Seven days after transplanting, the roots were drenched with a 75 mM sodium carbonate-sodium bicarbonate buffer solution (pH 9.0-10.0), and different concentrations (0, 1, 10, 100, 1000 ng / ml) of penicillin solution were added. The plant height of the common wheat was recorded after 30 days of treatment.

[0072] The results are as follows Figure 3 As shown in Figures A through E, different wheat varieties responded differently to different concentrations of *Penicillium wansii*, but all could effectively alleviate salt-alkali stress at 1000 ng / ml.

[0073] Example 4, Soil cultivation experiment: Wan's penicillin increases the yield of ordinary wheat.

[0074] Common wheat was cultured using the method described in Example 3.

[0075] (1) Overview of the test site and test materials The test site is located at Building B5, No. 3366 Longteng Road, Beijipo Street, Daiyue District, Tai'an City, Shandong Province.

[0076] Common wheat varieties: Jimai 22, B6-3-3, Yannong 999, and Zhongguochun.

[0077] Salt-alkali stress treatment: The salt-alkali stress treatment was to drench the roots of vernalized wheat seedlings with 50 ml of 75 mM sodium carbonate-sodium bicarbonate buffer (pH 9.0-10.0). Sodium carbonate and sodium bicarbonate were purchased from Sigma-Aladin (USA).

[0078] (2) Experimental Design Germination of common wheat seeds: Place common wheat seeds in a petri dish lined with filter paper, add an appropriate amount of water, and germinate at room temperature.

[0079] Jointed wheat seedlings subjected to 30 days of continuous low-temperature light treatment: After germination at room temperature for 2 days, the wheat seedlings were moved to a 4℃ low-temperature environment for 24-hour light treatment and vernalization treatment for 30 days.

[0080] Salt-alkali stress after transplanting: Vernalized common wheat seedlings were transplanted into plastic pots (9×9×10 cm) filled with nutrient soil, with 5 seedlings per pot. Seven days after transplanting, the roots were drenched with a 75 mM sodium carbonate-sodium bicarbonate buffer solution (pH 9.0-10.0) and a 1 μg / ml solution of penicillin. After 14 days of treatment, photos were taken and chlorophyll levels were recorded. After 35-45 days of treatment, the common wheat matured, and the grains were harvested. The grain length, width, and thousand-grain weight were recorded.

[0081] result: like Figure 4 As shown, treatment with 1 μg / mL of *Penicillium wani* significantly increased the chlorophyll content of three wheat varieties: Jimai 22, B6-3-3, and YN999, but had no significant effect on Chinese spring wheat.

[0082] Under salt-alkali stress, the chlorophyll content of Jimai 22, B6-3-3, YN999 and Chinese Spring was significantly reduced. Treatment with 1 μg / mL of Wan's penicillin could reverse the effect of salt-alkali stress on the chlorophyll content of B6-3-3 and YN999, but had almost no effect on Chinese Spring.

[0083] like Figure 5 As shown, compared with the controls of Jimai 22, B6-3-3, and Yannong 999, the application of 1 μg / ml of Wan's penicillin significantly increased the thousand-grain weight and grain width of Jimai 22. Figure 5 A); B6-3-3 and Yannong 999 both showed significant improvements in grain length and grain width traits ( Figure 5 (B, C). In summary, treatment with 1 μg / mL of *Penicillium wansii* significantly improved key grain traits such as thousand-grain weight, grain length, and grain width in different wheat varieties.

[0084] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. The application of Wan's penicillin in improving wheat's resistance to salt and alkali stress, characterized in that, Wheat seeds can be treated with penicillin by adding penicillin, or seedlings can be treated by drenching with penicillin.

2. The application of the penicillin mirabilis according to claim 1 in improving wheat resistance to salt and alkali stress, characterized in that, The concentration of Wan's penicillin added should not be less than 1 μg / ml.

3. The application of the penicillin mirabilis according to claim 1 in improving wheat resistance to salt and alkali stress, characterized in that, The salt-alkali stress condition was achieved by treatment with a sodium carbonate-sodium bicarbonate buffer solution.

4. The application of the penicillin mirabilis according to claim 1 in improving wheat resistance to salt and alkali stress, characterized in that, In the aforementioned salt-alkali stress conditions, the pH value of the sodium carbonate-sodium bicarbonate buffer solution is 9.0-10.

0.

5. The application of the penicillin mirabilis according to claim 1 in improving wheat's resistance to salt and alkali stress, characterized in that, The wheat is selected from jointed wheat and common wheat.

6. The application of the penicillin mirabilis according to claim 5 in improving wheat resistance to salt and alkali stress, characterized in that, The common wheat varieties mentioned are Jimai 22, B6-3-3, and Yannong 999.

7. The application of the penicillin mirabilis according to claim 6 in improving wheat resistance to salt and alkali stress, characterized in that, Applying 1 μg / ml of Wan's penicillin to B6-3-3 and Yannong 999 completely reversed salt-alkali stress.

8. The application of the penicillin mirabilis according to claim 6 in improving wheat resistance to salt and alkali stress, characterized in that, Applying penicillin to plants under saline-alkali stress conditions can promote the growth of B6-3-3 and Yannong 999.

9. The application of the penicillin mirabilis according to claim 6 in improving wheat resistance to salt and alkali stress, characterized in that, Applying penicillin wannabese under saline-alkali stress conditions can increase wheat yield, wherein the wheat is selected from Jimai 22, B6-3-3 and Yannong 999.

10. A method for planting wheat in saline-alkali land, characterized in that, Includes the following steps: S1, wheat seeds germinate at room temperature; S2, Low-temperature continuous light treatment of wheat seedlings: After germination at room temperature for 2-5 days, wheat seedlings are moved to a low-temperature environment of 2-7℃ for 24-hour light treatment and vernalization treatment for 30-35 days. S3, transplant the vernalized wheat seedlings into the soil and treat the roots with Wan's penicillin solution.

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