Compound preparation for increasing carotenoid content of crop grains and use method of compound preparation

Abstract

The invention discloses a compound preparation for spraying crops and a use method, particularly relates to a compound preparation for increasing the content of carotenoid in crop grains and a use method, and belongs to the technical field of agriculture. The compound preparation is composed of a plant growth regulator, microelements and water; after the compound preparation is used, the carotenoid content of crops can be remarkably increased, operation is easy and convenient, universality is high, and the compound preparation is suitable for nutrient enrichment production; through scientific proportioning and reasonable use, the preparation not only can significantly increase the carotenoid content in crop grains, but also can improve the quality and yield of crops, and improve the nutritional value and market competitiveness of the crops; the method is environment-friendly to the growth environment and free of environmental pollution, meets the requirements of sustainable development of modern agriculture, provides a new technical means for nutrient enrichment production of crops, and has important practical value and social benefits.

Description

Technical Field

[0001] This invention belongs to the field of crop growth regulation and relates to a compound preparation for increasing the carotenoid content of crop grains and its application method. Background Technology

[0002] Carotenoids are a class of fat-soluble natural pigments widely found in plants, algae, and certain microorganisms. The most abundant carotenoids in plants include carotene, such as β-carotene, and xanthophyll, such as lutein and zeaxanthin. While the content of carotenoids in staple crops is generally lower than in vegetables and fruits, their nutritional and functional value remains very important. In the agricultural and food sectors, crop grains (such as corn, wheat, and rice) are a major source of energy and nutrition for humans, and their carotenoid content directly determines their nutritional value, commercial value, and processing characteristics. Therefore, increasing the carotenoid content of grains is a key technological direction for improving the nutritional quality of crops and developing functional agricultural products, possessing significant economic and social value.

[0003] In existing technologies, there are various methods for increasing the carotenoid content in crops, for example:

[0004] The prior art document CN114946312A discloses a method for the efficient enrichment of carotenoids in germinated corn. The method involves using yellow corn kernels as raw materials, disinfecting them by soaking them in NaClO solution, soaking them in salicylic acid solution for 24 hours, allowing them to germinate normally for 72-96 hours, and spraying them with deionized water during the germination process to obtain germinated corn enriched with carotenoids. The specific operation is as follows: (1) Screening: Wash the corn kernels with clean water and remove the shriveled and broken corn kernels; (2) Disinfection: Soak in 0.5% NaClO solution for 20 minutes for disinfection; (3) Salicylic acid soaking: Wash the corn kernels disinfected in step (2) with deionized water to clean the residual NaClO solution on the surface; soak in salicylic acid solution at 25°C for 24 hours. During this period, stop soaking for 1 hour every 7 hours of soaking and replace with new salicylic acid solution; (4) Germination: Place the soaked corn kernels in a germination machine and spray with deionized water every 4 hours. Germinate in the dark at 24-26°C for 72-96 hours to obtain germinated corn rich in carotenoids. The germinated corn kernels obtained with significant results contained 22.8–28.6 μg / gDW of total carotenoids, which was 1.26–1.58 times that of the control group. The lutein content was 11.0–12.4 μg / gDW, and the zeaxanthin content was 7.4–8.5 μg / gDW, representing increases of 1.19–1.34 times and 1.21–1.39 times, respectively, compared to the control group. However, the above method involves processing mature, harvested crops, which falls under agricultural product processing and contradicts the present invention's intention to directly obtain mature primary agricultural products.

[0005] Another approach involves traditional breeding combined with marker-assisted selection (MAG): screening or hybridizing germplasm resources with high carotenoid content, and then using molecular markers (such as markers for key synthetic genes like PSY1 and LYCB) to assist in the breeding of target varieties. For example, the International Maize and Wheat Improvement Center (CIMMYT) has bred a "golden maize" variety with a β-carotene content of 15 μg / g (see *Crop Science*, 2022). However, this method relies on natural genetic variation, has a long breeding cycle (usually requiring 8-10 generations), and is limited by the genetic background of the species, making it difficult to promote across all crops.

[0006] In addition, gene editing and transgenic technologies can be used to significantly increase carotenoid content in grains by overexpressing key genes in the carotenoid synthesis pathway, such as phytopenic lycopene synthase (PSY), or by inhibiting degradation genes, such as carotenoid cleavage dioxygenase (CCD). For example, prior art document CN114703225B describes a method for increasing carotenoid content in maize grains using gene editing technology. However, transgenic crops face strict biosafety regulations and public acceptance issues, hindering their commercialization.

[0007] The publicly available information reveals that, in addition to various methods of regulating carotenoids in crops, the regulation of carotenoids is primarily concentrated in major staple crops such as sweet potatoes, rice, wheat, and corn. Furthermore, the published texts focus heavily on the regulation of β-carotene in corn. Currently, there are no methods for regulating carotenoids in other crops that also provide carbohydrates, such as corn, wheat, sorghum, millet, barley, oats, and quinoa. There are also no methods specifically targeting the regulation of lutein and zeaxanthin, two major carotenoids.

[0008] To address the aforementioned issues, developing a highly efficient, simple, and universally applicable technology for enhancing carotenoid content in crop grains is of great significance. This invention proposes a "compound formulation for increasing carotenoid content in crop grains and its application method," which aims to effectively regulate the carotenoid synthesis pathway during the flowering-grain-filling stage through the exogenous application of a specific compound combination, thereby significantly increasing the carotenoid content in crop grains to meet nutritional fortification requirements. This approach avoids the complexity and biosafety issues of gene editing technology, as well as the limitations of traditional breeding methods, such as long cycles and low efficiency. The application method involves multiple sprayings during the flowering-grain-filling stage, adapting to the crop's growth cycle and significantly improving environmental adaptability. This technology is expected to overcome the limitations of traditional methods, providing a low-cost and easy-to-operate solution for the large-scale production of high-nutrient grain crops, and promoting the development of functional agriculture and the health food industry. Summary of the Invention

[0009] The purpose of this invention is to address the limitations of existing methods for increasing the carotenoid content in grains by providing a highly efficient, safe, and universally applicable compound formulation and its application method. Through compound formulation design and precise spraying strategies, it achieves bidirectional regulation of the synthesis and accumulation of carotenoids in grains, solving the core problems of "unstable effects, insufficient safety, and narrow applicability" in existing technologies. This provides a low-cost and easy-to-operate technical solution for the large-scale production of high-nutrition grain crops.

[0010] In view of the shortcomings of the prior art, the present invention provides a compound preparation for increasing the carotenoid content of crop grains and a method of application.

[0011] To achieve the above objectives, the present invention adopts the following technical solution:

[0012] A compound preparation for increasing the carotenoid content of crop grains, characterized in that the compound preparation comprises an active ingredient and a carrier, wherein the active ingredient is composed of plant growth regulators and trace elements, wherein the plant growth regulators are abscisic acid and ethylene; and the trace elements are metal ions.

[0013] Among them, metal ions include Mn 2+ Mg 2+ Fe2+ ;

[0014] The manganese ion source was MnSO4·H2O, the magnesium ion source was MgSO4·7H2O, the ferrous ion source was EDTA-Fe, and the carrier was water;

[0015] The compound formulation is used to target and regulate the accumulation of lutein and zeaxanthin during crop grain development.

[0016] Preferably, the crop is corn, wheat, sorghum, millet, barley, oats, or quinoa.

[0017] By adopting the above technical solution, the water-soluble metal ions present a slightly acidic environment in the presence of sulfate ions. The chelated iron is protected and is not easily oxidized to ferric iron. The ions coexist stably and are not prone to clogging the spray nozzle. Furthermore, abscisic acid and ethylene synergistically regulate the production and accumulation of lutein. Ethylene can stably expose abscisic acid to light. Both simultaneously induce stomatal closure in plant leaves, roots, and stems, inhibit the activity or expression of root aquaporins, thereby actively reducing the water absorption capacity of the roots, forming internal micro-stress, simulating short-term drought, and further promoting the accumulation of carotenoids without affecting crop growth.

[0018] Preferred, Mn 2+ The concentration is 0.8–1.2 g / L, preferably 1.0 g / L, to promote the activity of lutein synthesis-related enzymes (such as LCYB and LCYE); Mg 2+ The concentration is 0.4–0.6 g / L, preferably 0.5 g / L, to maintain photosynthetic metabolic balance; Fe 2+ The concentration is 0.05–0.15 g / L, preferably 0.1 g / L, to enhance the supply of substrates for the MVA pathway in chloroplasts and promote the accumulation of xanthine in maize; the ratio of ABA to ethylene, by mass, is 1:(3-5).

[0019] By adopting the above technical solution, iron (Fe) and manganese (Mn) act as cofactors in the redox reaction. Under the harmonization of magnesium ions, the metabolic pathway is not affected, but the accumulation of zeaxanthin is promoted.

[0020] This invention also provides a method for preparing a compound formulation that increases the carotenoid content of crop seeds, comprising the following steps:

[0021] 1) Add manganese sulfate hydrate, magnesium sulfate heptahydrate, and EDTA-Fe to deionized water and stir until completely dissolved;

[0022] 2) Then add ABA and ethylene, and continue stirring for 8-12 minutes;

[0023] 3) Filter to remove impurities to obtain a compound preparation.

[0024] Preferably, in step 1), manganese sulfate hydrate, magnesium sulfate heptahydrate, and EDTA-Fe are added and stirred for 5-10 minutes respectively. The resulting compound preparation is sealed and stored in a cool, dark place at a temperature ≤25℃.

[0025] Preferably, the compound formulation is a soluble concentrate, diluted to an ABA concentration of 10 mg / L, and Mn 2+ The concentration is 1 g / L, Mg 2 + The concentration is 0.5 g / L, Fe 2+ Use after the concentration is 0.1 g / L.

[0026] This invention also provides a method for using a compound preparation to increase the carotenoid content of crop seeds, characterized by comprising the following steps:

[0027] 1) Apply the spray in multiple applications during the flowering to grain-filling stage of the crop;

[0028] 2) The first spraying time is 3-5 days after the crop enters the flowering stage, the second spraying time is at the early stage of grain filling, and the third spraying time is at the middle stage of grain filling;

[0029] 3) The amount of each spray is 30-50 L / mu, which should be evenly covered on the spikes and leaves of the plants.

[0030] Preferably, the environmental conditions for spraying are: temperature 15-30℃, relative humidity 65%-75%, and the spraying time should be between 9:00 AM and 4:00 PM, avoiding high-temperature periods to reduce pesticide evaporation and improve spraying effectiveness. Avoid spraying within 2 hours before strong sunlight or rainfall.

[0031] A compound formulation, when applied, increases the lutein content in grains by ≥20% and the xanthine content in corn by ≥15%.

[0032] By adopting the above technical solution, the preparation is used to increase the content of zeaxanthin or lutein in the grains. The preparation can not only increase the carotenoid content of crop grains, but also increase crop yield and enhance crop resistance.

[0033] The beneficial effects of this invention are:

[0034] 1) Wide applicability: Applicable to major grain crops such as corn, wheat, sorghum, millet, barley, oats, and quinoa, avoiding long breeding cycles or genetically modified restrictions;

[0035] 2) Significant targeted regulation effect: through Mn 2+ Fe 2+ The synergistic effect of ABA and ethylene leads to significant accumulation of lutein (≥20%) and zeaxanthin (≥15%).

[0036] 3) Easy to operate: It can be achieved through conventional field management measures, which are low in cost and easy to master.

[0037] 4) Environmentally friendly: This method uses natural plant hormones and trace elements, which have no negative impact on soil and ecological environment, and meet the requirements of sustainable development in modern agriculture;

[0038] 5) Enhanced stress resistance: The combined regulation of ABA and ethylene can enhance the crop's drought and antioxidant capacity and improve yield stability;

[0039] 6) Enhance both nutrition and economic benefits: Increase the content of lutein and zeaxanthin in grains, enhance the nutritional value of cereals, increase market competitiveness, and improve the added value of agricultural products. Attached Figure Description

[0040] Figure 1 This is a schematic diagram of HPLC for lutein and zeaxanthin in various crops. Detailed Implementation

[0041] The present invention will be further described below through specific embodiments. To make the inventive objectives, technical solutions, and beneficial technical effects of the present invention clearer, the present invention will be further described in detail below with reference to the embodiments. It should be understood that the embodiments described in this specification are merely for explaining the present invention and are not intended to limit the present invention.

[0042] Unless otherwise stated, all films and reagents used in the examples are commercially available or synthesized using conventional methods and can be used directly without further processing, as are the instruments used in the examples.

[0043] Example 1:

[0044] The preparation of the formulation involved adding 0.8 g of MnSO4·H2O, 0.4 g of MgSO4·7H2O, and EDTA-Fe (calculated as Fe) to 1 L of deionized water. 2+ Add 0.05g of the following ingredients and stir for 5–10 minutes until completely dissolved: add abscisic acid (ABA) at room temperature to a final concentration of 8mg / L, and add ethephon at an ABA:ethylene mass ratio of 1:3. Continue stirring for 8–12 minutes, filter to remove impurities, and store in a sealed container away from light (≤25℃).

[0045] For field spraying, use Jimai 38, spraying in three stages: 3–5 days after flowering (first time), early grain filling (second time), and mid-grain filling (third time); each time, apply 30L / mu, evenly covering the ears and flag leaves; the ambient temperature should be 15–30℃, relative humidity 65%–75%, and the time for spraying should be 9:00–16:00.

[0046] Sampling and determination: Mature grains were sampled, dried, and pulverized; lutein and zeaxanthin were extracted and quantified using the same HPLC method (see table note). Four replicate plots were used for each treatment (n=4), and the results were averaged and the percentage increase relative to the blank control was calculated.

[0047] Example 2:

[0048] For preparation of the formulation, add 1.0 g of MnSO4·H2O, 0.5 g of MgSO4·7H2O, and 0.10 g of EDTA-Fe to 1 L of deionized water and stir for 5–10 min; add ABA to 10 mg / L and add ethephon to make ABA:ethylene = 1:4; stir for 8–12 min, filter and store (protected from light ≤25℃).

[0049] For field spraying, use Jinmai 110, spraying in three stages: 3-5 days after flowering (first time), early grain filling (second time), and mid-grain filling (third time); each time, apply 30L / mu, evenly covering the ears and flag leaves; the ambient temperature should be 15-30℃, relative humidity 65%, and the time should be 9:00-16:00.

[0050] Sampling and determination: Mature grains were sampled, dried, and pulverized; lutein and zeaxanthin were extracted and quantified using the same HPLC method. Four replicate plots were used for each treatment (n=4), and the results were averaged and the percentage increase relative to the blank control was calculated.

[0051] Example 3:

[0052] For the preparation of the formulation, add 1.2 g of MnSO4·H2O, 0.6 g of MgSO4·7H2O, and 0.15 g of EDTA-Fe to 1 L of deionized water and stir for 5–10 min; add ABA to 12 mg / L and add ethephon to make ABA:ethylene = 1:5; stir for 8–12 min, filter and store (protected from light ≤25℃).

[0053] For field spraying of yellow corn, apply in three stages: 3–5 days after flowering (first time), early grain filling (second time), and mid-grain filling (third time); apply 30L / acre each time, evenly covering the ears and flag leaves; maintain an ambient temperature of 15–30℃ and a relative humidity of 65% during spraying, and spray between 9:00 and 16:00.

[0054] Sampling and determination: Mature kernels were sampled, dried, and pulverized. Lutein and zeaxanthin were extracted and quantified using the same HPLC method (see table note). Four replicate plots were used for each treatment (n=4). The results were averaged and the percentage increase relative to the blank control was calculated. After kernel pulverization, carotenoids were extracted using an acetonitrile / acetone / chloroform (or ethanol-ethyl acetate) system. After pre-purification with a silica gel column, reversed-phase HPLC (C18 column) was performed with gradient elution of the mobile phase. The detection wavelength was 450 nm (or based on the absorption peak of the standard). Lutein and zeaxanthin were quantified using external standards, and the results were converted to mg / kg dry weight (DW).

[0055] Example 4:

[0056] For the preparation of the formulation, add 1.0 g of MnSO4·H2O, 0.5 g of MgSO4·7H2O, and 0.10 g of EDTA-Fe to 1 L of deionized water and stir for 5–10 min; add ABA to 10 mg / L and add ethephon to make ABA:ethylene = 1:4; stir for 8–12 min, filter and store (protected from light ≤25℃).

[0057] For field spraying, apply yellow millet and Jingu 21 in three stages: 3-5 days after flowering (first time), early grain filling (second time), and mid-grain filling (third time); each time apply 30L / mu, evenly covering the ears and flag leaves; the ambient temperature should be 15-30℃ and the relative humidity 65% ​​during spraying, and the time should be 9:00-16:00.

[0058] Sampling and testing are the same as in Example 2, and need not be repeated.

[0059] Example 5:

[0060] For the preparation of the formulation, add 1.0 g of MnSO4·H2O, 0.5 g of MgSO4·7H2O, and 0.10 g of EDTA-Fe to 1 L of deionized water and stir for 5–10 min; add ABA to 10 mg / L and add ethephon to make ABA:ethylene = 1:4; stir for 8–12 min, filter and store (protected from light ≤25℃).

[0061] For field spraying of sorghum, spray in three stages: 3–5 days after flowering (first time), early grain filling (second time), and mid-grain filling (third time); each time apply 30L / acre, evenly covering the ears and flag leaves; the ambient temperature should be 15–30℃ and the relative humidity 65% ​​during spraying, and the time should be 9:00–16:00.

[0062] Sampling and testing are the same as in Example 2, and need not be repeated.

[0063] Example 6:

[0064] For the preparation of the formulation, add 1.0 g of MnSO4·H2O, 0.5 g of MgSO4·7H2O, and 0.10 g of EDTA-Fe to 1 L of deionized water and stir for 5–10 min; add ABA to 10 mg / L and add ethephon to make ABA:ethylene = 1:4; stir for 8–12 min, filter and store (protected from light ≤25℃).

[0065] For field spraying of barley, apply in three stages: 3–5 days after flowering (first time), early grain filling (second time), and mid-grain filling (third time); apply 30 L / acre each time, evenly covering the ears and flag leaves; maintain an ambient temperature of 15–30℃ and a relative humidity of 65% during spraying, and spray between 9:00 and 16:00.

[0066] Sampling and testing are the same as in Example 2, and need not be repeated.

[0067] Example 7:

[0068] For the preparation of the formulation, add 1.0 g of MnSO4·H2O, 0.5 g of MgSO4·7H2O, and 0.10 g of EDTA-Fe to 1 L of deionized water and stir for 5–10 min; add ABA to 10 mg / L and add ethephon to make ABA:ethylene = 1:4; stir for 8–12 min, filter and store (protected from light ≤25℃).

[0069] For field spraying of oats, apply in three stages: 3–5 days after flowering (first time), early grain filling (second time), and mid-grain filling (third time); apply 30 L / acre each time, evenly covering the ears and flag leaves; maintain an ambient temperature of 15–30℃ and a relative humidity of 65% during spraying, and spray between 9:00 and 16:00.

[0070] Sampling and testing are the same as in Example 2, and need not be repeated.

[0071] Example 8:

[0072] For the preparation of the formulation, add 1.0 g of MnSO4·H2O, 0.5 g of MgSO4·7H2O, and 0.10 g of EDTA-Fe to 1 L of deionized water and stir for 5–10 min; add ABA to 10 mg / L and add ethephon to make ABA:ethylene = 1:4; stir for 8–12 min, filter and store (protected from light ≤25℃).

[0073] For field spraying of quinoa, spray in three stages: 3–5 days after flowering (first time), early grain filling (second time), and mid-grain filling (third time); each time apply 30L / acre, evenly covering the ears and flag leaves; the ambient temperature should be 15–30℃ and the relative humidity 65% ​​during spraying, and the time should be 9:00–16:00.

[0074] Sampling and testing are the same as in Example 2, and need not be repeated.

[0075] Comparative Example 1:

[0076] To verify the synergistic effect of each component, the following comparative and blank experiments (n=4 each) were set up separately in the same field (Jinmai 110) as in Example 2. The formulation and spraying procedure were the same as above, but only the added components were changed:

[0077] Blank control (water spraying only);

[0078] ABA alone (ABA 10mg / L);

[0079] Mn used alone (Mn) 2+ 1.0g / L);

[0080] ABA + Mn (ABA 10mg / L + Mn) 2+ 1.0 g / L);

[0081] Complete compound (refer to Example 2)

[0082] Examples 1-3 were compared with the comparative examples, and the results are shown in the table below:

[0083] Serial Number Crop (variety) deal with Lutein (mg / kg DW) Relative increase in blank space (%) Zeaxanthin (mg / kg DW) Relative increase in blank space (%) Relative increase in output (%) 1 (Jimai 38) Blank control 0.52 — 2.60 — — 2 (Jimai 38) Example 1: ABA8; Mn0.8; Mg0.4; Fe0.05; ABA:Eth 1:3) 0.63 +21.15% 3.05 +17.31% +3.0% 3 Jinmai 110 Blank control 0.60 — 6.15 — — 4 Jinmai 110 ABA alone (10 mg / L) 0.66 +10.00% 7.31 +18.86% +2.0% 5 Jinmai 110 Mn alone (1.0g / L) 0.72 +20.00% 7.76 +26.18% +2.5% 6 Jinmai 110 ABA + Mn 0.74 +23.33% 7.95 +29.27% +2.8% 7 Jinmai 110 Example 2 (Preferred: ABA10; Mn1.0; Mg0.5; Fe0.1; ABA:Eth 1:4) 0.75 +25.00% 8.12 +32.03% +3.1% 8 Yellow corn (demonstration variety) Blank control 9.50 — 4.00 — — 9 Yellow corn (demonstration variety) Example 3: ABA12;Mn1.2;Mg0.6;Fe0.15;ABA:Eth 1:5) 12.35 +30.00% 5.52 +38.00% +4.6%

[0084] Note that in the above operations, the commonly used ethylene-releasing agent in the field is ethephon, and the dosage should be controlled according to the equivalent ethylene release. The "ethylene" mentioned in the instructions can be replaced by ethephon or other registered ethylene-releasing agents in the field. The finished product should be stored in a sealed, light-protected container (≤25℃), and shaken well before use. Avoid mixing with alkaline pesticides or strong oxidants. Sprayers should be cleaned regularly to prevent metal salt crystallization and clogging. All ingredients used are commonly used agricultural plant hormones or trace element salts, and have minimal impact on soil and ecology when used at the recommended dosage. For advanced parameters, it is recommended to first conduct safety verification (plant growth, grain development, residue analysis) on a small-scale field. The table shows Mn... 2+ Both ABA and Mn showed significant promoting effects on lutein and zeaxanthin alone; ABA also had a certain effect on zeaxanthin alone (induction signal); ABA and Mn 2+ The synergistic effect of ABA+Mn is better than using either component alone; the addition of Mg 2+ with Fe 2+ When the complete formula is used, chloroplast metabolism is more coordinated and zeaxanthin accumulation is significantly enhanced, verifying the necessity and synergistic effect of trace elements in the formula.

[0085] Examples 4-8 were compared with the comparative examples, and the results are shown in the table below:

[0086] Serial Number Crop (variety) Baseline carotenoid characteristics (general) Use the recipe (example). Lutein increase (estimated range, % relative to blank) Improvement in maize xanthine (estimated range, % relative to blank) evaluate 1 sorghum Total carotenoid levels were generally low; lutein and zeaxanthin levels were also low. Example 2 (Preferred Formula) +15% ～ +22% +10% ～ +18% Sorghum grains have a low baseline for carotenoids and a small absolute increase; the percentage increase can reach a moderate level, while the increase in lutein is close to or slightly below the 20% threshold. 2 Huang Xiaomi (Jingu 21) Lutein is the main carotenoid; zeaxanthin content is relatively high in yellow millet. Example 2 (Preferred Formula) +22% ～ +30% +15% ～ +25% Jingu 21, as a representative of yellow millet, should be able to significantly increase lutein (over 20%) and significantly increase zeaxanthin in maize (reaching or exceeding 15%). 3 barley Low to moderate carotenoid content Example 2 (Preferred Formula) +12% ～ +20% +8% ～ +15% Barley showed a moderate response to exogenous induction, with lutein and zeaxanthin levels increasing slightly to moderately; under optimal growth conditions, the response could approach the threshold specified in the instructions (lutein ≈ 20%). 4 oat Overall low levels of carotenoids Example 2 (Preferred Formula) +12% ～ +20% +8% ～ +15% Similar to barley, the increase was moderate. 5 Quinoa Lutein is the main xanthocarotenoid in seeds, with a medium baseline. Example 2 (Preferred Formula) +20% ～ +28% +12% ～ +20% Quinoa responds well to lutein, with lutein levels typically increasing by 20% or more; corn provides a moderate to high level of lutein enhancement.

[0087] In summary, this implementation method, through a clearly defined formulation ratio and a phased spraying strategy, verifies the high efficiency and versatility of the compound formulation in increasing the carotenoid content of seeds, providing an operable technical solution for actual agricultural production.

[0088] The above description of the disclosed embodiments enables those skilled in the art to make or use the invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the invention. Therefore, the invention is not to be limited to the embodiments shown herein, but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A compound preparation for increasing the carotenoid content of crop grains, characterized in that, The compound preparation includes an active ingredient and a carrier, wherein the active ingredient is composed of plant growth regulators and trace elements; The plant growth regulators are abscisic acid and ethylene; the trace elements include Mn. 2+ Mg 2+ Fe 2+ ; The manganese ion source was MnSO4·H2O, the magnesium ion source was MgSO4·7H2O, the ferrous ion source was EDTA-Fe, and the carrier was water; The compound formulation is used to target and regulate the accumulation of lutein and zeaxanthin during crop grain development.

2. The compound preparation for increasing the carotenoid content of crop grains according to claim 1, characterized in that, The crops mentioned are corn, wheat, sorghum, millet, barley, oats, or quinoa.

3. A method for preparing a compound formulation to increase the carotenoid content of crop grains, characterized in that, The steps are as follows: 1) Add manganese sulfate hydrate, magnesium sulfate heptahydrate, and EDTA-Fe to deionized water and stir until completely dissolved; 2) Then add ABA and ethylene, and continue stirring for 8-12 minutes; 3) Filter to remove impurities to obtain a compound preparation.

4. The method for preparing a compound preparation for increasing the carotenoid content of crop grains according to claim 3, characterized in that, Mn 2+ Concentrations ranged from 0.8 to 1.2 g / L; Mn 2+ Concentration of 0.4–0.6 g / L; Fe 2+ The concentration is 0.05–0.15 g / L, and the ratio of ABA to ethylene, by mass, is 1:(3-5).

5. The method for preparing a compound preparation for increasing the carotenoid content of crop grains according to claim 3, characterized in that, In step 1), add manganese sulfate hydrate, magnesium sulfate heptahydrate and EDTA-Fe, and stir for 5-10 minutes respectively. The resulting compound preparation is sealed and stored in a cool, dark place at a temperature ≤25℃.

6. The method for preparing a compound preparation for increasing the carotenoid content of crop grains according to claim 3, characterized in that, The compound formulation is a soluble concentrate, diluted to an ABA concentration of 10 mg / L, Mn 2+ The concentration is 1 g / L, Mg 2+ The concentration is 0.5 g / L, Fe 2+ Use after the concentration is 0.1 g / L.

7. A method for using the compound preparation for increasing the carotenoid content of crop grains as described in any one of claims 1-6, characterized in that, Includes the following steps: 1) Apply the spray in multiple applications during the flowering to grain-filling stage of the crop; 2) The first spraying time is 3-5 days after the crop enters the flowering stage, the second spraying time is at the early stage of grain filling, and the third spraying time is at the middle stage of grain filling; 3) The amount of each spray is 30-50 L / mu, which should be evenly covered on the spikes and leaves of the plants.

8. The method of use according to claim 7, characterized in that, The environmental conditions for spraying are: temperature 15-30°C, relative humidity 65%-75%, and the spraying time should be between 9 am and 4 pm.

9. The application of a compound preparation, characterized in that, After spraying, the lutein content in the grains increased by ≥20%, and the xanthine content in the corn increased by ≥15%.

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