Application of sakuranetin in improving drought resistance of wheat in seedling stage

Abstract

The invention discloses application of sakuranetin 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 application of the sakuranetin or the derivative thereof in improving the plant drought resistance or / and preparing the product for improving the plant drought resistance. The invention further provides a method for improving the drought resistance of plants, and the method comprises the step of applying sakuranetin to the plants so as to improve the drought resistance of the plants. According to the method, the seeds of a plurality of conventional wheat varieties are used as test materials, the influence of the exogenous sakuranetin on the survival rate and biomass in the wheat seedling stage under the drought stress is researched, the physiological mechanism of the externally applied sakuranetin in the wheat seedling stage under the drought stress is discussed, and a practical technology is provided for promoting how to improve the drought resistance in the seedling stage when the wheat is planted in a dry land in production.

Description

Technical Field

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

[0002] Wheat is one of the most important food crops in the world. Most of its planting areas are arid or semi-arid regions. 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. 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 aminoethyl hexanoate (DA-6), chlorfenapyr, sodium nitrophenolate, paclobutrazol, polyamines and other plant hormone substances, which can effectively improve crops to increase food production.

[0003] Sakurain (5,4'-dihydroxy-7-methoxyflavanone) belongs to the flavonoids and was first extracted and isolated from the branches of cherry trees. Later, the key enzyme gene in the synthesis of sakurain, sakurain synthase (naringenin-7-O-methyltransferase, NOMT), was also isolated and identified in rice. Sakurain has always been considered an important plant antiviral factor in rice and other plants, which can actively respond to a variety of adverse environmental factors, including pathogen invasion, ultraviolet radiation, jasmonic acid treatment, etc. 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] In order to solve the above technical problems, in a first aspect, the present invention provides the use of sakura glycosides or their derivatives in improving plant drought resistance and / or preparing products that improve plant drought resistance.

[0006] Furthermore, in the application, the improving the drought resistance of plants may be improving the drought resistance of plants at the seedling stage.

[0007] 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.

[0008] Furthermore, in the application, the product may be a drought-resistant preparation.

[0009] Furthermore, in the application, the active ingredients of the drought-resistant preparation include sakura vinifera and / or sakura vinifera derivatives.

[0010] Furthermore, in the application, the sakura derivative can be sakura salt, sakura glycoside, etc.

[0011] Furthermore, in the application, the drought-resistant preparation can improve the tolerance of plants to drought stress.

[0012] 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.

[0013] Furthermore, in the application, the drought-resistant preparation also includes other ingredients, such as auxiliary agents necessary in the processing technology.

[0014] Furthermore, in the application, the plant may be any of the following:

[0015] B1) Monocots;

[0016] B2) Gramineae;

[0017] B3) Triticum plants;

[0018] B4) Wheat (Triticum aestivum L.).

[0019] To solve the above technical problems, in a second aspect, the present invention provides a method for improving plant drought resistance, the method comprising applying sakura chlorophyll to plants to improve the drought resistance of the plants.

[0020] Furthermore, in the method described above, the application method can be mixing the sakura glycoside solution with soil, or irrigating the roots, or spraying the leaves, or making it into a seed coating agent to wrap the seeds.

[0021] Furthermore, in the method, the application concentration of the sakura glycoside solution can be 10 μM-20 μM.

[0022] Furthermore, in the method, the plant may be any 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 to increase the survival rate and / or biomass 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 the embodiments of the present invention, the wheat is selected from Chinese Spring, Stone 4185, Fielder, Chang 6878, and Drought Selection No. 10.

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

[0031] 1. The present invention is the first to reveal the role of sakura extract in improving plant drought resistance;

[0032] 2. The aboveground fresh weight and survival rate after rehydration of the sakura glycosides-treated group were significantly higher than those of the control group, indicating that sakura glycosides 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 The changing trend of soil water content during drought stress treatment with exogenous application of sakura phenol and water as control;

[0034] Figure 2 The wheat seedlings before drought treatment (7-day-old seedlings) and during drought treatment (10-day-old seedlings) with exogenous application of sakura alkaloids and water. The scale bar is 2 cm.

[0035] Figure 3 The effect of sakura alkaloids on drought stress tolerance in wheat seedlings;

[0036] Figure 4 This is the effect of sakura chlorophyll on the survival rate of wheat seedlings after drought and rehydration. The scale bar is 2 cm. DETAILED DESCRIPTION

[0037] 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.

[0038] 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.

[0039] The quantitative tests in the following examples were repeated three times unless otherwise specified, and the results were averaged.

[0040] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[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] 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.

[0043] The Fielder in the following embodiments is preserved in this laboratory and disclosed in the paper "Bai X, Zhan G, Tian S, Peng H, Cui X, Islam MA, Goher F, Ma Y, Kang Z, Xu ZS, Guo J. Transcription factor BZR2 activates chitinase Cht20.2 transcription to confer resistance to wheatstripe rust. Plant Physiol. 2021 Dec 4; 187 (4): 2749-2762. doi: 10.1093 / plphys / kiab383. PMID: 34618056; PMCID: PMC8644182." 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.

[0044] The long 6878 in the following embodiments 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 aPremature 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.

[0045] The Hanxuan No. 10 in the following examples was kindly donated by Jing Ruilian's research group at the Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, and was disclosed in the paper "Guo J, Hao C, Zhang Y, Zhang B, Cheng X, Qin L, Li T, Shi W, Chang X, Jing R, Yang W, Hu W, Zhang X, Cheng S. Association and Validation of Yield-Favored Alleles in Chinese Cultivars of Common Wheat (Triticum aestivum L.). PLoS One. 2015Jun 11; 10(6): e0130029.doi: 10.1371 / journal.pone.0130029. PMID: 26067129; PMCID: PMC4466017." The public can obtain the biological material from the applicant in accordance with the relevant provisions of the national biosafety. The obtained biological material is only used for repeating the experiments of the present invention and cannot be used for other purposes.

[0046] The CAS number of sakurain in the following example is 2957-21-3, the purity is 99%, and it is purchased from Solebow Company with the product number B29195-10mg.

[0047] 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.

[0048] Example 1: Sakura extract improves drought tolerance of wheat seedlings

[0049] 1. Experimental Design

[0050] Select plump wheat seeds (Chinese Spring, Shi 4185, Fielder, Chang 6878, and Hanxuan No. 10) with complete embryos and uniform size, disinfected with 75% ethanol for 10 minutes, rinsed with distilled water 5 times, disinfected with 3% sodium hypochlorite for 10 minutes, and rinsed repeatedly with distilled water. After soaking in pure water for 12 hours, select white seeds for the experiment. According to the experimental grouping, the nutrient soil with the same water content (Danish nutrient soil (PINDSTRUP, Denmark)) was mixed with pure water or different concentrations of cherry blossom solution (10μM and 20μM) at a weight to volume ratio of 1:2 (g:mL), and each blue culture box was filled with 180g of mixed wet soil, and the soil in the blue culture box was leveled. The white wheat seeds were evenly placed on the flat soil, and 5 varieties were planted in each flower pot at the same time, with 22 seeds for each variety. After sowing, each pot was covered with 30g of nutrient soil. After sowing and covering with soil, watering was stopped for drought treatment. All seedlings were placed in an incubator with a light duration of 16 hours, a temperature of 25°C, and a humidity of 70%. The effects of different concentrations of sakura phenol on drought resistance of wheat seedlings were observed. When there was a significant difference between the sakura phenol treatment group and the control group, samples were taken for the determination and statistical analysis of seedling physiological indicators.

[0051] The sakurain solution was prepared by dissolving sakurain in anhydrous ethanol and adjusting the sakurain solution to a target concentration (10 μM or 20 μM) with pure water.

[0052] 2. Experimental treatment

[0053] The optimal concentrations (10μM and 20μM) of sakura extract selected in the preliminary experiment were mixed with soil and then sown as the experimental treatment group, and distilled water was mixed with soil and then sown as the control group. This experiment designed 3 treatment groups, each treatment group was set up with three culture boxes for replication, and each culture box was divided into 22 seeds of each of Chinese Spring, Shi 4185, Fielder, Chang 6878, and Hanxuan No. 10.

[0054] Treatment group 1. Control group (control): pure water was mixed with soil, and watering was stopped after sowing to carry out drought treatment.

[0055] Treatment group 2. 10 μM sakura glycosides experimental group: 10 μM sakura glycosides solution was mixed with soil, and watering was stopped after sowing to carry out drought treatment.

[0056] Treatment group 3. 20 μM sakura glycosides experimental group: 20 μM sakura glycosides solution was mixed with soil, and watering was stopped after sowing to carry out drought treatment.

[0057] 3. Index determination

[0058] 1. Determination of soil moisture content

[0059] 1.1. Determination of soil moisture content after 10, 12 and 15 days of drought treatment

[0060] The control group and the sakura extract experimental group were both watered immediately after sowing and subjected to drought treatment. 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 culture box, and all soil samples were weighed using a 1 / 10,000 balance.

[0061] 1.2. Change trend of soil moisture content in different treatment groups during drought stress treatment

[0062] The experimental results are as follows Figure 1 As shown, Figure 1 The results of soil moisture measurement in the control group and two different sakura phenol concentration treatment groups are shown. The results show that the soil moisture content in both the control group and the experimental group gradually decreased with the change of drought days, but there was no significant difference in soil moisture content between the experimental group and the control group.

[0063] 2. Seedling growth

[0064] 2.1. Determination of seedling growth after 10 days of drought treatment

[0065] After 10 days of drought treatment, the wheat seedlings in the sakura glycosides experimental group and the control group were investigated and statistically analyzed, including the length of the aboveground part, the fresh weight (FW) of the aboveground part, the dry weight (DW) of the aboveground part, and the relative water content of the aboveground part. Three biological replicates were measured for each variety, and every 5 seedlings were one biological replicate.

[0066] 2.2 Effects of exogenous application of sakura extract on wheat seedlings under drought stress

[0067] Figure 2 The wheat seedlings before drought treatment (7-day-old seedlings) and during drought treatment (10-day-old seedlings) were treated with sakura extract and water. Figure 2 It showed that there was no significant difference between the control group and the externally applied sakura polyphenols at different concentrations before drought treatment. However, as the soil moisture content decreased during drought stress, the wheat seedlings in the experimental group with externally applied sakura polyphenols grew significantly better and taller than those in the control group.

[0068] Figure 3 To study the effect of exogenous application of sakura extract on wheat seedlings under drought stress. Figure 3 The statistical results of medium A showed that the external application of 20 μM sakura phenol significantly reduced the aboveground height of China Spring, while the reduction in aboveground plant height of other varieties (Shi 4185, Fielder, Chang 6878, and Hanxuan No. 10) did not reach a significant level. Figure 3Figures B and C show that exogenous application of sakura chlorophyll significantly increased the fresh weight (FW) and relative water content of the aboveground parts (FW-DW) / FW*100% of different varieties under drought stress conditions. Figure 3 Figure D shows that exogenous application of sakura phenol had no significant effect on the dry weight (DW) of wheat seedlings under drought stress.

[0069] 3. Survival rate after rehydration

[0070] 3.1. Survival rate statistics after rehydration

[0071] 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 in different treatment groups were counted.

[0072] 3.2 Effect of exogenous application of sakura extract on the survival rate of wheat seedlings after drought and rewatering

[0073] The results are as follows Figure 4 As shown, Figure 4 The effect of exogenous application of sakura glycosides on the survival rate of wheat seedlings after drought rehydration. The results showed that compared with the control group, the exogenous application of 10μM and 20μM sakura glycosides solution could significantly improve the survival rate of wheat seedlings after drought stress in China Spring, Shi 4185, Fielder, Chang 6878 and Hanxuan No. 10; especially for China Spring, the survival rate of the group treated with 10μM and 20μM sakura glycosides solution increased significantly by nearly one-fold compared with the control group. This shows that sakura glycosides mixed with soil can significantly improve the drought resistance of wheat seedlings of multiple different varieties, but there may be some differences in the effect of improving drought resistance of different wheat varieties.

[0074] 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. Application of sakura glycosides or their derivatives in improving plant drought resistance or / and preparing products that improve plant drought resistance.

2. The use according to claim 1, characterized in that: The improving of plant drought resistance is improving the drought resistance of plants at the seedling stage.

3. The use according to claim 1 or 2, 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.

4. The use according to any one of claims 1 to 3, characterized in that: The product is a drought-resistant preparation.

5. The use 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.).

6. A method for improving plant drought resistance, characterized in that: The method comprises applying sakura chlorophyll to plants to enhance the drought resistance of the plants.

7. The method according to claim 6, characterized in that: The application method is to mix the sakura solution with soil, or irrigate the roots, or spray it on the leaves, or make it into a seed coating agent to wrap the seeds.

8. The method according to claim 6 or 7, characterized in that: The sakurain solution is applied at a concentration of 10 μM to 20 μM.

9. The method according to any one of claims 6 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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