Biostimulant for promoting germination of wine grapes and application of biostimulant

Abstract

The invention discloses a biostimulant for promoting germination of wine grapes and application of the biostimulant, and belongs to the technical field of grape planting. The biostimulant comprises the following components: resveratrol, sodium alginate and chitin. The invention also provides a spraying liquid for promoting germination of wine grapes and a preparation method of the spraying liquid, the preparation method comprises the following steps: dissolving the resveratrol in ethanol to obtain a resveratrol ethanol solution, respectively dissolving the sodium alginate and the chitin in hot water to obtain a sodium alginate aqueous solution and a chitin aqueous solution, and mixing the sodium alginate aqueous solution with the chitin aqueous solution to obtain the spraying liquid for promoting germination of wine grapes. And finally, fully mixing the resveratrol ethanol solution, the sodium alginate aqueous solution and the chitin aqueous solution, and filtering to obtain the spraying liquid. The biostimulant provided by the invention can promote germination and growth of wine grapes and improve uniformity and robust degree of germination. After the biostimulant provided by the invention is used, the wine grapes can enter a germination stage earlier, and a solid foundation is laid for subsequent growth and development.

Description

Technical Field

[0001] The invention relates to the technical field of grape planting, in particular to a biostimulant for promoting budding of wine grapes and application thereof. Background Art

[0002] Grapes have evolved a winter dormancy strategy to resist the threat of cold. Winter bud dormancy is not only an effective biological characteristic for grape survival, but also a biological event that affects the economic efficiency of grape production. Dormancy and germination are two continuous biological processes. Bud dormancy is an adaptation mechanism of temperate deciduous fruit trees. During dormancy, plant growth is inhibited, cell division is blocked, and respiratory and metabolic activities slow down, allowing plants to adapt to seasonal environmental changes and survive in harsh winter environments. Only when the environment is suitable can the buds enter the germination state.

[0003] Resveratrol is a polyphenol compound commonly found in grape skins, red wine and other plants. It has antioxidant and anti-inflammatory effects. In agricultural applications, it is believed to enhance the stress resistance of crops, such as resistance to diseases and environmental stress. Currently, there are few studies on the use of resveratrol as a germination stimulant. Sodium alginate, also known as alginate or sodium alginate, is a natural polysaccharide extracted from seaweed. It is an oligomer formed by α-L-mannuronic acid and β-D-guluronic acid connected by glycosyl bonds. Alginate oligosaccharide (AOS) is a degradation product of sodium alginate with a degree of polymerization of 2 to 10. It has good water solubility and a wide range of biological activities. Studies have found that AOS can promote plant growth, enhance plant resistance to cold, and alleviate stress damage to plants. Chitin is a natural polysaccharide extracted from the shells of crustaceans and is also a type of biostimulant. It can promote the development of plant roots, enhance plant immunity, and help resist pathogens in the soil. However, there is relatively little research on resveratrol, sodium alginate and chitosan in promoting germination of wine grapes, and the germination-promoting effect and mechanism of action in the grape system are still unclear. Summary of the invention

[0004] The purpose of the present invention is to provide a biostimulant for promoting budding of wine grapes and its application, so as to solve the problems existing in the above-mentioned prior art. The present invention uses resveratrol, sodium alginate and chitosan in combination to play a synergistic effect.

[0005] To achieve the above object, the present invention provides the following solutions:

[0006] Technical Solution 1: A biostimulant for promoting germination of wine grapes, comprising resveratrol.

[0007] Furthermore, the components also include sodium alginate and chitosan.

[0008] Furthermore, the mass fraction of the resveratrol in the biostimulant is 45-55 parts by mass.

[0009] Furthermore, in terms of mass fraction, the mass fraction of the sodium alginate in the biostimulant is 15-35 parts.

[0010] Furthermore, the mass fraction of chitosan in the biostimulant is 15-25 parts by mass.

[0011] Technical Solution 2: A spray liquid for promoting budding of wine grapes, the active ingredient of which includes the biostimulant.

[0012] Technical Solution 3: The method for preparing the spray liquid comprises first dissolving the resveratrol in ethanol to obtain a resveratrol ethanol solution, then dissolving the sodium alginate and the chitosan in hot water respectively to obtain a sodium alginate aqueous solution and a chitosan aqueous solution, and finally fully mixing the resveratrol ethanol solution, the sodium alginate aqueous solution and the chitosan aqueous solution, and filtering to obtain the spray liquid.

[0013] Technical Solution 4: Application of the biostimulant in promoting grape budding.

[0014] Furthermore, the biostimulant is sprayed during the grape budding period.

[0015] The present invention discloses the following technical effects:

[0016] The present invention provides a biostimulant for promoting the germination of wine grapes, comprising the following ingredients: resveratrol, sodium alginate and chitin. Among them, resveratrol can stimulate the physiological activity inside the grape plant, enhance the division and growth ability of cells, and provide sufficient power for germination; sodium alginate can improve the soil structure, increase the water and fertilizer retention capacity of the soil, create a good soil environment for grape germination, and ensure that the root system can better absorb nutrients and water; chitin is a natural polysaccharide substance with good biocompatibility and biodegradability. In wine grapes, chitin can induce grapes to produce resistance reactions, improve the immunity and stress resistance of grapes, and reduce the occurrence of diseases and insect pests. The present invention uses resveratrol, sodium alginate and chitin in combination, which can play a synergistic role, better promote the germination and growth of wine grapes, and improve the uniformity and robustness of germination. After using the biostimulant, wine grapes can enter the germination period earlier, laying a solid foundation for subsequent growth and development.

[0017] The present invention uses wine grape branches as test materials, studies the germination and development of grape winter buds under the treatment of the biostimulant of the present invention, and detects hydrogen peroxide (H2O2), superoxide anion (OFR), superoxide dismutase (SOD), malondialdehyde (MDA), plant starch, fructose (FT), soluble sugar and total protein content of the winter buds under different treatments, clarifies the correlation between the winter bud germination process and physiological and biochemical characteristics, and provides a basis for the formulation of grape winter bud germination control measures. DETAILED DESCRIPTION

[0018] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0019] It should be understood that the terms described in the present invention are only for describing a particular embodiment and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. The intermediate value in any stated value or stated range, and each smaller range between any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0020] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0021] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.

[0022] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0023] Example 1

[0024] 1 Materials and Methods

[0025] 1.1 Materials and reagents

[0026] The test material Chardonnay was planted in Ningxia Yongning Grape Technology Courtyard.

[0027] 1.2 Biostimulant treatment

[0028] Collect annual branches with uniform growth and no disease, select strong and plump winter buds, cut the upper incision flatly and the lower incision obliquely to prepare single bud cuttings.

[0029] Preparation of biostimulant spraying liquid: first dissolve resveratrol in ethanol to obtain resveratrol ethanol solution, then dissolve sodium alginate and chitosan in 60°C hot water respectively to obtain sodium alginate aqueous solution and chitosan aqueous solution, and finally, according to the weight parts, respectively: 50 parts of resveratrol ethanol solution, 25 parts of sodium alginate aqueous solution and 20 parts of chitosan aqueous solution are fully mixed and filtered to obtain spraying liquid 1; 55 parts of resveratrol ethanol solution, 35 parts of sodium alginate aqueous solution and 25 parts of chitosan aqueous solution are mixed and filtered to obtain spraying liquid 1. The liquid was fully mixed and filtered to obtain spray liquid 2; 45 parts of resveratrol ethanol solution, 15 parts of sodium alginate aqueous solution and 15 parts of chitosan aqueous solution were fully mixed and filtered to obtain spray liquid 3; 40 parts of resveratrol ethanol solution, 40 parts of sodium alginate aqueous solution and 30 parts of chitosan aqueous solution were fully mixed and filtered to obtain spray liquid 4; the resveratrol ethanol solution was filtered to obtain spray liquid 5; 50 parts of sodium alginate aqueous solution and 45 parts of chitosan aqueous solution were fully mixed and filtered to obtain spray liquid 6;

[0030] On the day of collection, spray solution 1-6 and water control were sprayed on winter buds (0d), for a total of 7 groups of treatments, with water spraying as control. The branches were inserted into the sand and watered regularly to keep the water level slightly above the sand. They were cultured in a 25℃ long-day (16h / 8h) light incubator, and the germination rate was observed. The development of winter buds was observed every 6d. Winter buds were collected at 0, 6, 12, and 18d after treatment in each treatment group, and the branches and excess tissues were cut off around the winter buds. After quick freezing with liquid nitrogen, they were stored in a -80℃ refrigerator.

[0031] 1.3 Investigation of germination rate and observation of winter bud germination process

[0032] The buds were considered to have germinated when the winter bud scales cracked and the green tissue was clearly visible. The germination rate was calculated as a percentage. Three to four single bud cuttings were randomly selected from each group at 0, 6, and 12 days after the spraying treatment (the 18-day buds had already germinated to the stage of young leaves, so they were not observed). The front and side views of the winter buds were observed using a stereoscope (Leica Microsystems Co., Ltd., M205FCA model), and the structural changes of the cross and longitudinal sections of the winter buds were observed after manual sectioning.

[0033] 1.4 Physiological data The samples were crushed with liquid nitrogen using a medium-throughput tissue grinder (Beijing Dinghaoyuan Technology Co., Ltd., TL-2010S). Three biological replicates were set for the winter bud samples of each treatment group. Before the formal determination, 2 to 3 samples with large expected differences were selected for pre-determination. The 18-day samples had germinated into new shoots, so the relevant physiological indicators were not tested and were only used to calculate the germination rate. The kits for hydrogen peroxide, superoxide anion, superoxide dismutase, malondialdehyde, plant starch, fructose, plant soluble sugars and biuret protein content determination were purchased from Suzhou Keming Biotechnology Co., Ltd. The detection steps and numerical calculation methods refer to the kit instructions.

[0034] 1.5 Data processing Three biological replicates were set for the determination of each physiological index.

[0035] 2 Results and analysis

[0036] 2.1 Effects of spraying solutions 1-6 and water control on the germination rate of winter buds of grapes

[0037] The germination rates of grape winter buds under different spraying liquid treatments were counted. The results showed that compared with the clear water control, spraying liquids 1-4 promoted germination, and the germination rates of spraying liquids 5 and 6 were significantly lower than those of spraying liquids 1-4. At 18 days, except for the clear water control treatment group and spraying liquids 5 and 6, the germination rates of the spraying liquids 1-4 treatment groups exceeded 90%.

[0038] 2.2 Observation of winter bud germination process using stereomicroscope

[0039] The development of winter buds of grapes at different stages was observed under a stereomicroscope. At 12 days, the bud scales broke open, and the yellow hairs on the top of the winter buds were clearly visible. Except for the water control group and the spraying liquid 5 and 6 treatment groups, the green tissues of the winter buds could not be seen breaking out of the scales. In the spraying liquid 1-4 treatment groups, green tissues were visible, the hairs covering the winter buds were broken, and the length of the internodes on the cut surface increased. Among them, the winter buds in the spraying liquid 1 treatment group germinated most vigorously.

[0040] 2.3 Changes of physiological parameters during grape winter bud germination

[0041] 2.3.1 Changes in hydrogen peroxide and superoxide anion content

[0042] H2O2 and OFR are the most common reactive oxygen molecules in organisms. With the development of grape winter buds, the H2O2 content first decreased and then increased. There was no significant difference in H2O2 content under each spraying solution treatment at 6 days; at 12 days, the H2O2 content in the spraying solution 1-4 treatment groups was high, while the content of spraying solution 5 and 6 increased slowly, and the H2O2 content in the clear water control group was low. With the development of grape winter buds, at 6 days, the OFR content in the spraying solution 5 and 6 treatment groups was significantly lower than that in the clear water control group; at 12 days, except for the spraying solution 5 and 6 treatment groups, the OFR content in other treatment groups was significantly higher than that in the control group.

[0043] 2.3.2 Changes in SOD enzyme activity and MDA content

[0044] SOD can catalyze superoxide anion free radicals to generate oxygen and hydrogen peroxide in organisms, and plays a vital role in the balance of oxidation and anti-oxidation in the body. The SOD enzyme activity was detected during the germination of winter buds, and it was found that its content gradually increased with the development of the buds. Compared with the control, there was no significant difference in enzyme activity under each spraying liquid treatment at 6 days. The enzyme activity under spraying liquid 5 and 6 treatments was lower than that of the clear water control treatment group, and the enzyme activity of the other treatments was higher than that of the control; at 12 days, the winter bud enzyme activity under spraying liquid 1-4 treatments was significantly higher than that of the control, and the enzyme activity under the other spraying liquid treatment groups was not significantly different.

[0045] MDA is one of the important products of membrane lipid peroxidation. The level of lipid oxidation can be detected by detecting the MDA level. With the development of winter buds, the MDA content showed an overall downward trend, but the decline was not large, indicating that the degree of membrane lipid peroxidation was low or the ability to remove MDA was strong. The results showed that there was no significant difference in the MDA content among the treatment groups.

[0046] 2.3.3 Changes in carbohydrate and protein content

[0047] As the winter buds develop, the starch content of the plants decreases. Compared with the control group, the starch content of the winter buds treated with all the spraying liquid treatment groups was higher than that of the control group. Plant soluble sugars include glucose, fructose, sucrose, etc. During the period from dormancy to germination, plant starch can be converted into soluble sugars to provide available energy for plant growth. The present invention also detected the changes of fructose during dormancy to germination. As the winter buds of grapes develop, the fructose content in the winter buds first decreases and then increases. At 6 days, the fructose content of each treatment group reaches the lowest value, and the difference in fructose content among the groups is not significant; at 12 days, the fructose content increases, among which the fructose content of the spraying liquid 1-4 treatment group is the highest, and the fructose content of the spraying liquid 6 treatment group is the lowest. As the winter buds of grapes develop, the plant soluble sugar content in the winter buds shows a downward trend. At 12 days, the plant soluble sugar content of the spraying liquid 1-6 treatment groups is higher than that of the clear water control group. The soluble protein content can reflect the metabolic level of the plant. The general trend of soluble protein content in winter buds of each group showed a decrease first and then an increase, with the lowest content at 6 days. With the end of dormancy, the soluble protein content increased. At 6 days, the soluble protein content of the spraying liquid 1-6 treatment groups was lower than that of the control group. At 12 days, the protein content of the spraying liquid 5 and 6 treatment groups was lower than that of the control group, and the protein content of the other treatments was higher than that of the control group.

[0048] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. A biostimulant for promoting budding of wine grapes, characterized in that: Ingredients include resveratrol.

2. The biostimulant according to claim 1, characterized in that The components also include sodium alginate and chitosan.

3. The biostimulant according to claim 1, characterized in that Calculated by weight, the weight percentage of the resveratrol in the biostimulant is 45-55 parts.

4. The biostimulant according to claim 2, characterized in that Calculated by weight, the weight percentage of the sodium alginate in the biostimulant is 15-35 parts.

5. The biostimulant according to claim 2, characterized in that: Calculated by weight, the weight percentage of chitosan in the biostimulant is 15-25 parts.

6. A spray liquid for promoting budding of wine grapes, characterized in that: The active ingredient comprises the biostimulant according to any one of claims 1 to 5.

7. The method for preparing the spray liquid according to claim 6, characterized in that: Firstly, the resveratrol is dissolved in ethanol to obtain a resveratrol ethanol solution, and then the sodium alginate and the chitosan are respectively dissolved in hot water to obtain a sodium alginate aqueous solution and a chitosan aqueous solution. Finally, the resveratrol ethanol solution, the sodium alginate aqueous solution and the chitosan aqueous solution are fully mixed and filtered to obtain the spray solution.

8. Use of the biostimulant according to any one of claims 1 to 5 in promoting grape budding.

9. The use according to claim 8, characterized in that: The biostimulant is sprayed during the grape budding period.