Application of vitexin to improvement of drought resistance of wheat in seedling stage

Abstract

The invention discloses application of vitexin to improvement of drought resistance of wheat at a seedling stage, and belongs to the field of application of compounds. The technical problem to be solved by the invention is how to improve the stress resistance of plants. In order to solve the technical problem, the invention provides the method for improving the drought resistance of the plants, and the method comprises the step of applying vitexin to the plants so as to improve the drought resistance of the plants. The invention also provides application of vitexin or the derivative thereof in improving plant drought resistance or / and preparing a product for improving plant drought resistance. The method comprises the following steps: by taking seeds of a plurality of conventional wheat varieties as test materials, researching the influence of exogenous vitexin on the survival rate, fresh weight and relative water content in the seedling stage of wheat under drought stress, and discussing the physiological mechanism of externally applied vitexin in the seedling stage of wheat under drought stress; and a practical technology is provided for improving the drought resistance of the wheat in the seedling stage when the wheat is planted in the dry land during production.

Description

Technical Field

[0001] The invention belongs to the application field of compounds, and particularly relates to the application of vitexin in improving drought resistance of wheat at seedling stage. Background Art

[0002] Applying small molecules to improve plant tolerance to stress factors is an important way for many agricultural scientists to explore how to deal with crop yield reduction. They have developed a variety of different types of growth regulators, including DA-6, chlorfenapyr, sodium nitrophenolate, paclobutrazol, polyamines and other plant hormones, which can effectively improve crops to increase food production. Wheat is one of the most important food crops in the world. Most of its planting areas are arid or semi-arid areas. Drought is the most serious environmental factor causing crop yield reduction. Therefore, the application and research of improving wheat drought resistance can provide a solution to the food crisis.

[0003] Vitexin, also known as vitexin, belongs to the flavonoids and can actively respond to a variety of adverse environmental factors. Vitexin is a natural plant flavonoid glycoside compound extracted from the leaves of Vitex quinata, a plant of the Verbenaceae family. It is widely distributed in the leaves and stems of dozens of plants in nature, such as Vitex quinata, Vitex rotundifolia, Abacus, Crataegus pinnatifida, Ficus microcarpa, Alsophila spinulosa, Lygodium japonica, Fern, Broussonetia papyrifera, Rhododendron chinense, Folium isatidis, etc. The main sources are the leaves, stems and hawthorn of Vitex quinata plants of the Verbenaceae family. Summary of the invention

[0004] The technical problem to be solved by the present invention is: how to improve the stress resistance of plants, specifically, how to improve the drought resistance of wheat.

[0005] To solve the above technical problems, in a first aspect, the present invention provides a method for improving drought resistance of plants, the method comprising applying vitexin to plants to improve the drought resistance of the plants.

[0006] Furthermore, in the method described above, the application method can be mixing the aqueous solution of vitexin with soil, or irrigating the roots, or spraying the leaves, or making it into a seed coating agent to wrap the seeds.

[0007] Furthermore, in the method, the administration concentration of the vitexin aqueous solution may be 10 μM-20 μM.

[0008] Furthermore, in the method, the plant is any one of the following:

[0009] B1) Monocots;

[0010] B2) Gramineae;

[0011] B3) Triticum plants;

[0012] B4) Wheat (Triticum aestivum L.).

[0013] In order to solve the above technical problems, in a second aspect, the present invention provides the use of vitexin or its derivatives in improving plant drought resistance and / or preparing products for improving plant drought resistance.

[0014] Furthermore, in the application, the improving the drought resistance of plants may be improving the drought resistance of plants at the seedling stage.

[0015] Furthermore, in the application, the improvement of plant drought resistance may be to increase the survival rate of plants under drought stress or / and increase the fresh weight of plants under drought stress or / and increase the relative water content of plants under drought stress.

[0016] Furthermore, in the application, the product may be a drought-resistant preparation.

[0017] Furthermore, in the application, the active ingredients of the drought-resistant preparation include vitexin and / or vitexin derivatives.

[0018] Furthermore, in the application, the vitexin derivative can be vitexin salt, vitexin glucoside, vitexin rhamnoside, etc.

[0019] Furthermore, in the application, the drought-resistant preparation can improve the tolerance of plants to drought stress.

[0020] Furthermore, in the application, the drought-resistant preparation may be in various dosage forms, including but not limited to liquid, emulsion, suspension, powder, granule, wettable powder or water-dispersible granule, etc.

[0021] Furthermore, in the application, the drought-resistant preparation also includes other ingredients, such as auxiliary agents necessary in the processing technology.

[0022] Furthermore, in the application, the plant is any one of the following:

[0023] B1) Monocots;

[0024] B2) Gramineae;

[0025] B3) Triticum plants;

[0026] B4) Wheat (Triticum aestivum L.).

[0027] In the present invention, the improvement of plant drought resistance may specifically be increasing the biomass and / or survival rate of plants under drought stress.

[0028] In the present invention, the increasing of plant fresh weight may specifically be increasing the fresh weight of the above-ground part and / or the fresh weight of the underground part of the plant under drought stress.

[0029] In an embodiment of the present invention, the wheat is selected from Chang 6878, Shi 4185, Chinese Spring, and Shi 18.

[0030] The beneficial technical effects achieved by the present invention are as follows:

[0031] 1. The present invention discloses for the first time the role of vitexin in improving plant drought resistance;

[0032] 2. The fresh weight of the aboveground part and the survival rate after rehydration in the vitexin-treated group were significantly higher than those in the control group, indicating that vitexin can effectively improve the drought resistance of wheat at the seedling stage, providing theoretical guidance and scientific basis for the production and application of crops in arid fields. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 This is the changing trend of soil water content during drought stress treatment with exogenous application of vitexin and water as control.

[0034] Figure 2 This shows the growth of wheat seedlings after drought stress treatment with exogenous application of vitexin and water as control.

[0035] Figure 3 Exogenous application of vitexin significantly improves the drought resistance of wheat. DETAILED DESCRIPTION

[0036] The present invention is further described in detail below in conjunction with specific embodiments, and the examples provided are only for illustrating the present invention, rather than for limiting the scope of the present invention. The examples provided below can be used as a guide for further improvements by those of ordinary skill in the art, and do not constitute a limitation of the present invention in any way.

[0037] The experimental methods in the following examples, unless otherwise specified, are all conventional methods, and are performed according to the techniques or conditions described in the literature in the field or according to the product instructions. The materials, reagents, etc. used in the following examples, unless otherwise specified, can all be obtained from commercial channels.

[0038] The quantitative tests in the following examples were repeated three times unless otherwise specified, and the results were averaged.

[0039] The long 6878 in the following examples was kindly donated by Sun Daizhen's research group of the College of Agriculture of Shanxi Agricultural University, and was disclosed in the document "Lu J, Sun L, Jin X, Islam MA, Guo F, Tang X, Zhao K, Hao H, Li N, Zhang W, Shi Y, Wang S, Sun D. Analysis of physiological and transcriptomic differences between a premature senescence mutant (GSm) and its wild-type in common wheat (Triticum aestivum L.). Biology (Basel). 2022Jun 12; 11(6): 904. doi: 10.3390 / biology11060904. PMID: 35741425; PMCID: PMC9219967." The public can obtain the biological material from the applicant in accordance with the relevant provisions of national biosafety. The above-mentioned biological material is only used for repeating the experiments of the present invention and cannot be used for other purposes.

[0040] The Shi 4185 in the following examples was preserved in our laboratory and was used in the paper "Zhou Y, Liu J, Guo J, Wang Y, Ji H, Chu X, Xiao K, Qi X, Hu L, Li H, Hu M, Tang W, Yan J, Yan H, Bai X, Ge L, Lyu M, Chen J, Xu Z, Chen M, Ma Y. GmTDN1 improves wheat yields by inducing dual tolerance to both drought and low-N stress. Plant Biotechnol J. 2022Aug;20(8):1606-1621.doi:10.1111 / pbi.13836.Epub 2022May 25.PMID:35514029;PMCID:PMC9342622." The public may obtain the biological material from the applicant in accordance with the relevant provisions of national biosafety. The biological material obtained is only used for repeating the experiments of the present invention and cannot be used for other purposes.

[0041] The Chinese spring in the following examples is preserved in our laboratory and disclosed in the paper "Xu ZS, Liu L, Ni ZY, Liu P, Chen M, Li LC, Chen YF, Ma YZ. W55a encodes a novel protein kinase that is involved in multiple stress responses. J Integr Plant Biol. 2009 Jan; 51(1): 58-66. doi: 10.1111 / j.1744-7909.2008.00776.x. PMID: 19166495." The public can obtain the biological material from the applicant in accordance with the relevant national biosafety regulations. The obtained biological material is only used for repeating the experiments of the present invention and cannot be used for other purposes.

[0042] Shi 18 in the following embodiments is preserved in this laboratory and disclosed in the paper "Shi H, Chen M, Gao L, Wang Y, BaiY, Yan H, Xu C, Zhou Y, Xu Z, Chen J, Tang W, Wang S, Shi Y, Wu Y, Sun D, ​​Jia J, MaY. Genome-wide association study of agronomic traits related to nitrogen useefficiency in wheat. Theor Appl Genet. 2022Dec;135(12):4289-4302.doi:10.1007 / s00122-022-04218-5. Epub 2022Sep 22.PMID:36136127." The public can obtain the biological material from the applicant in accordance with the relevant provisions of the national biosafety. The above-mentioned biological material is only used for repeating the experiments of the present invention and cannot be used for other purposes.

[0043] The CAS number of vitexin in the following examples is 3681-93-4, the purity is 99%, and it was purchased from Solebao Company with the product number SV8180-20mg.

[0044] Statistical analysis of the experimental data was performed using Microsoft Excel 2016 for raw data processing, SPSS statistics 26.0 analysis software for one-way analysis of variance, and the Duncan method for multiple comparisons and significant difference tests. The significant difference was P < 0.05. Different letters in the figure indicate statistically significant differences. Graphpad Prism 8.0 was used for drawing.

[0045] Example 1: Application of vitexin in improving drought tolerance of wheat seedlings

[0046] 1. Experimental Design

[0047] Select plump wheat seeds (Chang 6878, Shi 4185, Chinese Spring, Shi 18) with complete embryos and uniform size, disinfect them with 75% ethanol for 10 minutes, rinse them with distilled water 5 times, disinfect them with 3% sodium hypochlorite for 10 minutes, and rinse them repeatedly with distilled water. After soaking in pure water for 12 hours, select the white seeds for the test. Dry nutrient soil with the same water content and water or different concentrations of vitexin solution (10μM and 20μM) are mixed at a weight to volume ratio of 1:2 (g:mL). Each round flower pot is filled with 240g of mixed wet soil and the soil is leveled. The white wheat seeds are evenly placed on the flat soil, 25 seeds are sown in each flower pot, and each pot is covered with 60g of dry nutrient soil after sowing. After sowing and covering with soil, watering was stopped for drought treatment in the control group and different concentrations of vitexin (10μM and 20μM), and all seedlings were placed in an incubator with a light duration of 16h, a temperature of 25°C, and a humidity of 70% to observe the effects of different concentrations of vitexin on drought resistance of wheat seedlings. Photos were taken when there was a significant difference between the 10μM and 20μM vitexin experimental groups and the control group.

[0048] The preparation method of the vitexin solution is as follows: dissolving vitexin in anhydrous ethanol and adjusting the concentration of vitexin to a target concentration (10 μM or 20 μM) with water.

[0049] 2. Experimental treatment

[0050] The optimal concentration (20 μM) of vitexin screened in the preliminary experiment was mixed with soil and then sown as the experimental treatment group, and distilled water mixed with soil was used as the control group. This experiment designed 3 treatment groups, each treatment group was set up with 3 pots of replication, and each flower pot was divided and sown with 25 seeds of Chang 6878, Shi 4185, Chinese Spring, and Shi 18.

[0051] 1. Control group: Mix soil with pure water, and stop watering after sowing to carry out drought treatment.

[0052] 2.10μM vitexin experimental group: 10μM vitexin solution was mixed with soil, and watering was stopped after sowing to carry out drought treatment.

[0053] 3.20μM vitexin experimental group: 20μM vitexin solution was mixed with soil, and watering was stopped after sowing to carry out drought treatment.

[0054] 3. Index determination

[0055] 1. Determination of soil moisture content after 10, 12 and 15 days of drought treatment

[0056] The control group and the vitexin experimental group were watered immediately after sowing. Soil samples were taken and weighed at 10, 12 and 15 days of drought treatment (W1), and then dried in an oven at 65°C for 5 days and weighed again (W2). The soil moisture content is (W1-W2) / W1*100%. Two soil samples were taken from each flower pot, and all soil samples were weighed with a 1 / 10,000 balance.

[0057] The results are as follows Figure 1 As shown, Figure 1 The changing trends of soil moisture content in different treatment groups during drought stress treatment. Figure 1 The results showed that at 10, 12 and 15 days of drought treatment, the soil moisture content in the treatment groups with different concentrations of vitexin was not significantly lower than that in the control group, but the soil moisture content decreased significantly during the drought stress process.

[0058] 2. Determine the growth of seedlings after 10 days of drought treatment

[0059] After 10 days of drought treatment, the wheat seedlings in the vitexin experimental group and the control group were investigated and statistically analyzed. The fresh weight of the aboveground part (FW) and the relative water content of the aboveground part (FW-DW) / FW*100% were measured. DW is the dry weight of the seedlings after weighing the fresh weight (FW) and drying them in a 65℃ oven for 5 days and then weighing them. Three biological replicates were measured for each variety, and each biological replicate had 5 seedlings.

[0060] Seedling growth Figure 2 As shown, the results showed that compared with the control (a) and (d), the application of 10μM (b) and (e) and 20μM (c) and (f) vitexin significantly slowed down the lodging and wilting of some wheat (Chang 6878, Shi 18) and improved the drought resistance of wheat ( Figure 2 ).

[0061] The results of the aboveground fresh weight and relative water content of the aboveground parts are as follows: Figure 3 As shown in (c) and (d), the results showed that: the exogenous application of 20 μM vitexin can significantly increase the fresh weight of the aboveground part of wheat (Chang 6878, Shi 4185, and China Spring), the exogenous application of 10 μM and 20 μM vitexin can significantly increase the relative water content of the aboveground part of some wheat (Chang 6878 and Shi 4185), and the relative water content of the aboveground part of some wheat (China Spring and Shi 18) has no significant difference with the control group, which indicates that the effect of vitexin on improving wheat drought resistance may be variety-dependent.

[0062] 3. Statistics of survival rate after rehydration

[0063] After 15 days of drought treatment, the soil moisture content dropped to about 45%. Each culture box was watered with 300 mL of water for rehydration. One week after rehydration, the survival rates of different varieties (Chang 6878, Shi 4185, China Spring, Shi 18) in different treatment groups were counted.

[0064] The results are as follows Figure 3 As shown in (a) and (b), the results showed that the application of 10 μM and 20 μM vitexin significantly increased the survival rate of wheat varieties Chang 6878, Shi 4185 and Shi 18, and the effect of 20 μM vitexin was better than that of 10 μM vitexin ( Figure 3 (a) and (b)).

[0065] The present invention has been described in detail above. For those skilled in the art, without departing from the purpose and scope of the present invention, and without the need to carry out unnecessary experimental conditions, the present invention can be implemented in a wide range under equivalent parameters, concentrations and conditions. Although the present invention provides specific embodiments, it should be understood that the present invention can be further improved. In a word, according to the principles of the present invention, the application is intended to include any changes, uses or improvements to the present invention, including departure from the disclosed scope in the application, and changes made with conventional techniques known in the art.

Claims

1. A method for improving plant drought resistance, characterized in that: The method comprises applying vitexin to a plant to increase the drought resistance of the plant.

2. The method according to claim 2, characterized in that: The application method is to mix the aqueous solution of vitexin with soil, or to irrigate the roots, or to spray the leaves, or to make a seed coating agent to wrap the seeds.

3. The method according to claim 1 or 2, characterized in that: The administration concentration of the vitexin aqueous solution is 10 μM-20 μM.

4. The method according to any one of claims 1 to 4, characterized in that: The plant is any of the following: B1) Monocots; B2) Gramineae; B3) Triticum plants; B4) Wheat (Triticum aestivum L.).

5. Use of vitexin or its derivatives in improving plant drought resistance or / and in preparing products for improving plant drought resistance.

6. The use according to claim 5, characterized in that: The improving of plant drought resistance is improving the drought resistance of plants at the seedling stage.

7. The use according to claim 5 or 6, characterized in that: The improving of plant drought resistance is to improve the survival rate of plants under drought stress or / and to improve the fresh weight of plants under drought stress or / and to improve the relative water content of plants under drought stress.

8. The use according to any one of claims 5 to 7, characterized in that: The product is a drought-resistant preparation.

9. The use according to any one of claims 5 to 8, characterized in that: The plant is any of the following: B1) Monocots; B2) Gramineae; B3) Triticum plants; B4) Wheat (Triticum aestivum L.).

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