Composite agent for promoting germination of grapes and use method of composite agent

Abstract

The invention discloses a combination agent for promoting grape germination and a use method thereof. The combined agent comprises one or more of lime sulfur, boric fertilizer, ethephon and sodium abietate. The treatment of the combination of lime sulfur and boron fertilizer accelerates the nutrient backflow, enhances the accumulation of branch dry matter and starch, effectively improves the germination speed, germination uniformity and flowering rate, and is successfully applied to the production of primary fruits of grape in season cultivation and secondary fruits of grape in staggered season cultivation for the first time. According to the method, nutrient accumulation of grape vines can be improved, germination is effectively promoted, technical support is provided for staggered season cultivation of grapes based on secondary fruits, and multiple positive effects of lime sulfur and boric fertilizer composite treatment on the sunshine rose grapes are disclosed for the first time. Particularly, the combined treatment of the lime sulfur and the boric fertilizer can be used as an innovative technology, and nutrient accumulation and flower bud differentiation in staggered season cultivation can be effectively promoted while the germination quality of the grapes under the conventional cultivation condition is improved.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of grape cultivation, and in particular, relates to a combined agent for promoting grape sprouting and a use method thereof. BACKGROUND

[0002] Grape (Vitis) is one of the oldest fruit trees in the world with a long history of cultivation. At present, the grape cultivation area in China ranks third in the world, and the yield ranks first (OIV Statistical Brief: Wine, Table Grapes & Dried Grapes in 2024, www.oiv.int). Dormancy is a typical feature of deciduous fruit trees, which is a biological adaptation of plants to changes in the external environment and seasonal changes, and is a periodic period in the process of plant growth and development (LANG GA. Dormancy: A new universal terminology [J]. Hort-Science, 1987, 22(5): 817-820). As a typical deciduous fruit tree, the sprouting uniformity of grape is closely related to the yield (Guo Songtao. Research on the effect of cold requirement and heat requirement and light supplement on grape sprouting in Ningxia greenhouse[D]. Ningxia University, 2020). However, in the southern warm winter regions such as Zhejiang, Yunnan, Hunan and Hainan, the temperature is generally high, and the accumulation of cold requirement is insufficient, which leads to inconsistent and irregular sprouting of grape, and further causes more time-consuming and labor-intensive problems in winter pruning and flower management in the peak flowering period. In the early and late forcing cultivation modes of grape (including the cultivation mode based on secondary fruit), forced defoliation leads to the delay of nutrient return, which seriously affects the rate and uniformity of subsequent sprouting. Studies have shown that the accumulation level of carbohydrates (such as starch) and mineral nutrients (such as nitrogen, potassium, boron, etc.) stored in the branches of perennial deciduous plants during the dormancy period directly affects the uniformity and speed of their sprouting (Zheng Yuanqing. Grape production period regulation technology in Zhangzhou area [J]. Acta Agronomica Sinica, 2024, 14(12): 50-54.). The differentiation and sprouting of flower buds in the next year of fruit trees cannot be separated from the efficient transport and storage of photosynthetic products by leaves before defoliation (Vivin P H, Castelan M, Gaudillère J P. A source / sink model to simulate seasonal allocation of carbon in grapevine [C] / / VI International Symposium on Computer Modelling in Fruit Research and Orchard Management 584. 2001: 43-56.).

[0003] In recent years, exogenous regulation measures are increasingly widely used in grape cultivation. Exogenous calcium ions can effectively break bud dormancy. Calcium signaling is involved in grape bud dormancy release, and HC and low temperature-induced dormancy release both depend on calcium signaling, which promotes grape bud germination by activating calcium-dependent protein phosphorylation and regulating gene expression (Pang X, Halaly T, Crane O, Keilin T, Keren-Keiserman A, Ogrodovitch A, Galbraith D, Or E. Involvement of calcium signalling in dormancy release of grape buds. J Exp Bot. 2007;58(12):3249-62. doi: 10.1093 / jxb / erm172. PMID: 17977848). Stone sulfur belongs to alkaline agents, which can not only stress leaves to promote leaf nutrient backflow, but also has a unique calcium ion component. In addition, boron, as a micronutrient, participates in sugar transport and cell wall formation, and can also synergize with calcium (Ca) to further optimize the germination process. Ethrel accelerates leaf senescence by inducing ethylene synthesis and has been widely used in fruit and vegetable trees, but its field application is significantly affected by climate and weather conditions such as temperature and humidity. Improper treatment can lead to reduced yield or even complete loss, and its effect on germination is controversial. Other treatments such as urea and potassium dihydrogen phosphate are also used to promote grape leaf fall and improve tree nutrition, thereby improving germination quality. SUMMARY

[0004] Based on this, the present application takes the sunlight rose grape of Hunan Agricultural University Sunlight Vineyard as the test material, sprays different combinations of stone sulfur, boron fertilizer, ethrel, etc. on the whole plant foliage, and systematically analyzes their effects on branch dry matter, starch content, germination speed and flowering rate. The present application determines the optimal grape nutrient regulation scheme; among the various treatment combinations, the best treatment is found to effectively promote grape germination speed and uniformity; and provides a theoretical basis and practical guidance for high-quality and efficient grape cultivation. The present application provides a theoretical basis and technical support for promoting nutrient backflow, promoting germination, and improving yield and quality in grape production.

[0005] Firstly, the present application provides a combination agent for promoting grape germination, which comprises one or more of stone sulfur, boron fertilizer, ethrel, and sodium abietate.

[0006] The combination agent preferably consists of the stone sulfur mixture and boron fertilizer; preferably, in the combination agent, the stone sulfur mixture is 10-20 times of 29% stone sulfur mixture, the boron fertilizer is 1g / L-4g / L; more preferably, the stone sulfur mixture is 15 times of 29% stone sulfur mixture, and the boron fertilizer is 2g / L.

[0007] In the combination agent, one of the stone sulfur mixture, the boron fertilizer, sodium abietate, or ethephon, ethephon+boron fertilizer, ethephon and urea+boron fertilizer can also be included; preferably, the stone sulfur mixture is 10-20 times of 29% stone sulfur mixture, the boron fertilizer is 1g / L-4g / L, the sodium abietate is 60g / L, and the ethephon is 600 times of 40% ethephon.

[0008] The application also provides the use of the combination agent in promoting grape sprouting.

[0009] The application also provides the method for promoting grape sprouting by the combination agent, which is to spray the combination agent on the whole grape plant in early November (more than 15 days before leaf fall in the normal season cultivation mode), early April or / and early May (more than 30 days before sprouting in the secondary fruit out-of-season cultivation mode).

[0010] The method preferably sprays the combination agent on the whole grape plant more than 15 days before leaf fall in the normal season cultivation mode or more than 30 days before sprouting in the secondary fruit out-of-season cultivation mode, and the combination agent is the stone sulfur mixture+boron fertilizer, wherein the stone sulfur mixture is 10-20 times of 29% stone sulfur mixture, and the boron fertilizer is 1g / L-4g / L.

[0011] The method more preferably the stone sulfur mixture is 15 times of 29% stone sulfur mixture, and the boron fertilizer is 2g / L.

[0012] Preferably, the grape is the Sunlight Rose grape.

[0013] The present application aims at the problems of slow sprouting speed, irregular sprouting, and difficulty in flowering in off-season cultivation of grape in southern regions. With Sunshine Rose grape as the test material, the physiological regulation effects of different reagents and composite treatments were innovatively explored. The effects of each combination treatment on branch nutrient accumulation, sprouting dynamics, and flowering ability under the conditions of one-time fruit conventional cultivation and two-time fruit off-season cultivation were systematically analyzed by whole plant spraying. The results showed that the treatment based on ethephon or urea was unstable, the main bud was severely necrotic, the flowering rate was low, and drug poisoning was easily induced. The treatment of stone sulfur mixture combined with boron fertilizer could promote leaf nutrient backflow, enhance branch dry matter and starch accumulation, effectively improve the sprouting speed, sprouting uniformity, and flowering rate. The combination showed the best sprouting promotion effect in conventional cultivation and was successfully applied to two-time fruit production in off-season cultivation for the first time. The key application window was determined to be the first half of May, and the best ratio was 15 times of stone sulfur mixture + 2 g / L of boron fertilizer, which was applied to whole plant spraying + shortening and leaf removal. The present application provides a relatively safe and efficient solution for difficult sprouting and irregular sprouting in southern regions, and provides technical support for off-season cultivation of grape based on two-time fruit.

[0014] The present application first discloses the multiple positive effects of stone sulfur mixture combined with boron fertilizer on Sunshine Rose grape: the combination agent and bud promotion method of the present application not only significantly promote leaf nutrient backflow and branch nutrient accumulation, but also effectively improve the sprouting speed and uniformity. What is particularly innovative is that the present application successfully applies the treatment scheme containing stone sulfur mixture and boron fertilizer to the production practice of two-time fruit of Sunshine Rose grape under off-season cultivation conditions for the first time. The results show that the combination treatment of stone sulfur mixture and boron fertilizer can be used as an innovative technology to improve the sprouting quality of grape under conventional cultivation conditions, and more effectively promote nutrient accumulation and flower bud differentiation in off-season cultivation. BRIEF DESCRIPTION OF DRAWINGS

[0015] Figure 1 Tree situation after 15 days of different treatments under normal season cultivation.

[0016] Figure 2 Dry weight proportion of one-year-old branches after different treatments under normal season cultivation.

[0017] Figure 3 Starch content proportion of one-year-old branches after 15 days of different treatments under normal season cultivation. Different lowercase letters indicate significant differences (P<0.05) between treatments.

[0018] Figure 4 Sprouting state at the same time under different treatments under normal season cultivation.

[0019] Figure 5 Sprouting rate under different treatments under normal season cultivation.

[0020] Figure 6The sprouting rate of the grapevine under the different treatments in the main season. Different lowercase letters represent significant differences between treatments (P<0.05).

[0021] Figure 7 The flowering rate of the grapevine under the different treatments in the main season. Different lowercase letters represent significant differences between treatments (P<0.05).

[0022] Figure 8 The branch status of the grapevine under the different treatments after 15 days in the off-season.

[0023] Figure 9 The sprouting rate of the grapevine under the different treatments in the off-season.

[0024] Figure 10 The main bud necrosis rate of the grapevine under the different treatments in the off-season. Different lowercase letters represent significant differences between treatments (P<0.05).

[0025] Figure 11 The main bud necrosis rate of the grapevine under the different treatments in the off-season. Different lowercase letters represent significant differences between treatments (P<0.05).

[0026] Figure 12 The starch content of the grapevine under the different treatments after 15 days in the off-season. Different lowercase letters represent significant differences between treatments (P<0.05).

[0027] Figure 13 The sprouting rate of the grapevine under the different treatments in the off-season.

[0028] Figure 14 The sprouting rate of the grapevine under the different treatments in the off-season.

[0029] Figure 15 The sprouting rate of the grapevine under the different treatments in the off-season. Different lowercase letters represent significant differences between treatments (P<0.05).

[0030] Figure 16 The flowering rate of the grapevine under the different treatments in the off-season. Different lowercase letters represent significant differences between treatments (P<0.05). DETAILED DESCRIPTION

[0031] In order to make the technical problems, technical solutions and beneficial effects to be solved by the present application more clear and explicit, the present application is described in detail and completely below in combination with embodiments. It should be noted that the specific embodiments described herein are only used to explain the present application and are not used to limit the present application. Obviously, the described embodiments are only a part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor fall within the scope of protection of the present application.

[0032] The experimental methods used in the following examples are conventional methods unless otherwise specified. The materials, reagents, instruments and equipment used in the following examples can be obtained from commercial channels unless otherwise specified.

[0033] 1. Materials and methods

[0034] 1.1 Test materials

[0035] The experiment was carried out from April 2024 to April 2025. The material was 8-year-old flybird-shaped trellis Sunlight Rose grape plants planted in the rain-sheltered cultivation base of Hunan Agricultural University in Changsha, Hunan Province (113°196′E, 28°132′N), with a row spacing of 2.8 m x 1.8 m, and about 45 main shoots were retained for each tree.

[0036] The control and the effective concentration of the plant growth regulator treatment were: 40% ethephon (600 times), potassium dihydrogen phosphate (100 g / L), 29% lime sulfur (10 times, 15 times, 20 times), urea (100 g / L), sodium abietate (60 g / L), boron fertilizer (1 g / L, 2 g / L, 4 g / L), water and leaf removal. Each reagent combination is shown in 1.2. The reagents used were all Guoguang brand, purchased from Sichuan Runer Technology Co., Ltd.

[0037] 1.2 Test method

[0038] For the regular season cultivation, Sunlight Rose grape trees with similar tree vigor were selected. On November 5, 2024, when the treatment was sprayed, the tree was in the stage of 50 days after fruit harvest, about 30 days before winter yellowing and leaf fall, and basically in the stage of growth stop.

[0039] Test one: each treatment and control was repeated 3 trees. 29% lime sulfur, sodium abietate, 40% ethephon, 40% ethephon + boron fertilizer, 29% lime sulfur + boron fertilizer, urea + boron fertilizer, urea, water and leaf removal were used for whole plant spraying, a total of 9 combinations.

[0040] For the production of secondary fruit in off-season cultivation, to explore the regulation of each treatment on grape sprouting under off-season cultivation mode, Sunlight Rose grape with consistent growth was selected. After removing the first flower, the effects of different treatments on sprouting speed and uniformity were determined. From April to June 2024, different reagent combinations (test two) and different concentrations of the best combination (test three) were treated in different test areas. At this time, Sunlight Rose grape trees were about 40 days after the sprouting period.

[0041] Test two, design each treatment and control 3 trees each. 29% lime sulfur, 29% lime sulfur + boron fertilizer, urea, urea + boron fertilizer, sodium abietate, sodium abietate + boron fertilizer, 40% ethylene, 40% ethylene + boron fertilizer, and water were used for whole plant spraying.

[0042] Test three, design each treatment and control 3 trees each. Based on the results of the previous pre-test, 29% lime sulfur (10 times) + boron fertilizer (1 g / L), 29% lime sulfur (10 times) + boron fertilizer (2 g / L), 29% lime sulfur (10 times) + boron fertilizer (4 g / L), 29% lime sulfur (15 times) + boron fertilizer (1 g / L), 29% lime sulfur (15 times) + boron fertilizer (2 g / L), 29% lime sulfur (20 times) + boron fertilizer (1 g / L), 29% lime sulfur (20 times) + boron fertilizer (2 g / L), 29% lime sulfur (20 times) + boron fertilizer (4 g / L), and water were used for whole plant spraying.

[0043] Test four, to determine the best treatment time for the optimal combination in the production of secondary fruit in off-season cultivation, design each treatment and control 3 trees each. On April 25, 2024, May 5, 2024, May 25, 2024, and June 25, 2024, 29% lime sulfur (15 times) + boron fertilizer (2 g / L) was used for whole plant spraying.

[0044] 1.2.1 Dry weight ratio detection

[0045] Leaf yellowing and shedding were observed 15 days after spraying. 2-3 node mid 0.5 cm stem samples were taken from branches with consistent growth 15 days after treatment, labeled, weighed quickly with an electronic balance, then placed in an oven at 105°C for 30 minutes, then adjusted to 80°C and dried to constant weight. The weight was taken every certain time until the difference between two weighings was less than 0.01g. Finally, the dry weight ratio was calculated by dividing the dry weight by the fresh weight. Dry weight ratio (%) = (DW / FW) x 100%.

[0046] 1.2.2 Starch content analysis

[0047] Starch content in the shoots was determined by anthrone method (Hendgen, M; Günther, S; Schubert, S; Löhnertz, O. Ethephon Activates the Transcription of Senescence-Associated Genes and Nitrogen Mobilization in Grapevine Leaves (Vitis vinifera cv. Riesling). Plants 2021, 10, 333.). Briefly, the dried grape shoot samples were ground into powder with a mortar, and 0.1-0.2 g of the powder was accurately weighed into a centrifuge tube after passing through a 60-mesh sieve. 10 mL of 80% ethanol was added, and the mixture was shaken several times in a boiling water bath for 30 minutes. After cooling, the mixture was centrifuged at 4000 rpm for 10 minutes, and the supernatant was discarded to remove soluble sugars. 10 mL of distilled water was added to the precipitate, and the mixture was heated in a boiling water bath for 30 minutes to fully dissolve the starch. After cooling, the mixture was centrifuged again, and the supernatant was collected and diluted to 50 mL in a volumetric flask. 1 mL of the diluted solution was taken into a test tube, 5 mL of anthrone reagent was added, and the mixture was colored in a boiling water bath for 10 minutes. After cooling, the absorbance was measured at 620 nm. At the same time, a standard curve was drawn using a glucose standard solution, and the starch content was calculated from the absorbance according to the standard curve. The result was expressed as mg of starch per g of dry sample.

[0048] 1.2.3 Statistics of main shoot necrosis rate

[0049] The main shoot necrosis was observed 15 days after the spraying treatment, and multiple shoots were randomly selected from the grape plants at the same time 15 days after the treatment. The main shoot necrosis was observed with a pair of tweezers, and the number of main shoot necrosis was recorded according to the standard of main shoot browning and drying. After the statistics were completed, the main shoot necrosis rate of each treatment group was calculated as follows: main shoot necrosis rate (%) = (number of main shoot necrosis / total number of shoots) x 100%.

[0050] 1.2.4 Statistics of germination rate

[0051] From the beginning of germination, the germination of each treatment group was observed regularly, and the number of winter shoots in each treatment was recorded according to the standard of bud swelling and green tissue exposure. After the germination period ended, the germination rate of each treatment group was calculated as follows: germination rate (%) = (number of germinated bud eyes / total number of bud eyes) x 100%.

[0052] 1.2.5 Statistics of germination rate

[0053] Starting from the early stage of germination, the number of germinations in each treatment group was counted every 2 or 3 days (based on the greening of the buds) until the end of the germination period; the germination rate at each time point was calculated as "germination rate (%) = (number of germinating buds / total number of buds) × 100%", and the mean and standard deviation of the repeated data for each group were calculated.

[0054] 1.2.6 Flowering Rate Statistics

[0055] After flowering, the flowering rate of each treatment group was observed and counted. The flowering status of each treatment group was observed regularly. Flowering was defined as the formation of obvious inflorescence primordia on buds or the appearance of complete inflorescences on branches. The number of branches or buds that formed inflorescences in each treatment replicate was counted, and the flowering rate of each replicate was calculated as "Flowering rate (%) = (Number of new shoots with flower spikes / Number of winter buds that have sprouted) × 100%".

[0056] 1.2.7 Data Statistical Analysis

[0057] One-way ANOVA was used to test the significant differences between different treatments, and Excel was used to generate graphs.

[0058] 2 Results and Analysis

[0059] 2.1 Effects of different treatments on bud break and flowering in primary grape production

[0060] 2.1.1 Effects of different treatments on the dry weight of one-year-old grape shoots after primary fruiting

[0061] Observe the changes in the Sunshine Rose grapevines on the 15th day after treatment (November 20th). Figure 1 The dry and wet weights of one-year-old grape branches were measured. The results (Table 1) showed significant differences in the percentage of dry weight among different treatments. The percentage of dry matter decreased in the leaf-removal treatment compared to before treatment, indicating that premature leaf removal not only hinders nutrient accumulation in the branches but also depletes them. The highest percentage of dry matter was achieved in the lime-sulfur + boron fertilizer combination at 49.09%, indicating that this treatment was beneficial for the accumulation of dry matter in the branches. Overall, the dry weight ratio of the lime-sulfur treatment group was generally higher than other combinations, while the effect of the water treatment was weaker. The addition of boron fertilizer had a significant effect on increasing dry matter; for example, compared to lime-sulfur treatment, the percentage of dry weight increased by 1.5% (…). Figure 2 ).

[0062] Table 1. Dry weight, starch content, germination rate, and flowering rate under different treatments.

[0063]

[0064] 2.1.2 Effects of different treatments on starch content in one-year-old branches of grapevines bearing primary fruit

[0065] The starch content in branches was determined using the anthrone method. The results showed significant differences in starch content among different treatments of *Sunshine Rose* (Table 1). The lime-sulfur + boron fertilizer treatment group had the highest starch content with minimal fluctuation, significantly better than the water treatment and leaf removal treatment. The starch content was significantly increased compared to before treatment, indicating that this combination was most conducive to starch accumulation. Lime-sulfur and sodium rosinate showed the next best results. The ethephon + boron fertilizer, ethephon, and urea + boron fertilizer treatments had moderate starch contents, but ethephon showed greater fluctuation. Urea and water treatments were relatively less effective, with the water treatment showing low and significantly fluctuating starch content. The leaf removal treatment had the lowest starch content, lower than the starch content of the branches before treatment, indicating that premature leaf removal not only hinders starch accumulation in the branches but also depletes it. Overall, treatments containing boron fertilizer or lime-sulfur have a significant effect on increasing the starch content of branches. Figure 3 ).

[0066] 2.1.3 Effects of different treatments on the bud break rate of one-year-old shoots of grapevines with primary fruit

[0067] From March 26th to April 10th, the germination rate of the terminal buds in each treatment group was recorded every 3 days. The germination rate of each treatment and the control group was compared with the dynamic data of germination speed (Table 2) and the germination status on the same day (Table 2). Figure 4 From the perspective of [data missing], there were significant differences in the germination process among different treatments. The lime-sulfur mixture + boron fertilizer showed the most outstanding performance and remained stable throughout the process, significantly higher than other treatments and the clean water group. Figure 5 The budding speed was slower with water and urea treatments. The leaf-removal treatment was significantly worse, with slow budding initiation and progress, significantly lower than other groups and the water treatment. Data indicates that foliar treatments that promote leaf yellowing and shedding before autumn leaf fall effectively enhance nutrient reserves in the branches, thus significantly increasing budding speed in the following spring. Among these, the lime-sulfur mixture and lime-sulfur mixture + boron fertilizer treatments showed the most outstanding performance, thanks to the synergistic effect of calcium ions in lime-sulfur mixture and boron fertilizer: calcium ions directly promote budding, while boron further enhances budding speed by increasing nutrient transport efficiency. Experimental results show that, under the same treatment conditions, adding boron fertilizer can further increase budding speed, confirming the positive regulatory effect of calcium and boron synergy on grape budding.

[0068] Table 2. Sprouting rate of winter buds under different treatments (unit: % / d)

[0069]

[0070] 2.1.4 Effects of different treatments on the bud break rate of one-year-old shoots of grapevines with primary fruiting

[0071] The germination rate of each treatment and control on April 12 was counted (Table 1). The results showed that the stone sulfur mixture and the stone sulfur mixture + boron fertilizer reached more than 90%, and the effect of urea treatment was weaker than other treatments, and was unstable, while the germination rate of water treatment and leaf removal was the worst, and each treatment was significantly different from water treatment and leaf removal. Overall, it is shown that spraying photos of promoting leaf yellowing and senescence on the leaves of Sunshine Rose grape before leaf fall in autumn can significantly promote the germination rate of grape in the second spring ( Figure 6 ).

[0072] 2.1.5 Effect of different treatments on the flowering rate of one-year-old branches of grape

[0073] The flowering rate of each treatment and control on April 23 was counted (Table 1). The results showed that compared with leaf removal treatment and water treatment, different treatments could improve the flowering rate of Sunshine Rose, and there was a significant difference. The flowering rate of leaf removal treatment was significantly lower than that of water treatment, indicating that early leaf removal treatment was not conducive to the differentiation of flower buds of Sunshine Rose grape. Sodium abietate, urea + boron fertilizer, ethylene + boron fertilizer, ethylene, and urea showed moderate performance in promoting the flowering rate of Sunshine Rose grape, although there was no significant difference between them, but there was a significant difference between them and water treatment and leaf removal treatment; it is shown that the above treatments can promote the differentiation of flower buds of Sunshine Rose grape to different degrees, but the stability of urea is poor. The flowering rate of stone sulfur mixture + boron fertilizer treatment reached more than 94%, and there was a significant difference between it and other treatments, water control and leaf removal control. Overall, it is shown that spraying stone sulfur mixture + boron on the leaves of Sunshine Rose grape before leaf fall in autumn can significantly promote the flowering rate of grape in the second spring ( Figure 7 ).

[0074] 2.2 Effect of different treatments on the germination and flowering of grape in the production of secondary fruit

[0075] 2.2.1 Effect of different treatments on the winter bud and main bud necrosis of main shoots

[0076] On May 20, the changes of grape branches after each treatment were observed ( Figure 8 ), and the main bud necrosis rate of each treatment and control and the main bud necrosis rate of stone sulfur mixture + boron fertilizer at different times were counted. From the column chart ( Figure 10) and statistical data (Table 3) Results found that the effects of different treatments on the main bud necrosis rate were significantly different. The main bud necrosis rate of the combination treatment group of calcium sulfate and boron was lower, and the standard deviation was smaller, indicating that its effect was more stable. The main bud necrosis rate of the urea treatment group was significantly higher than that of other treatments, and the standard deviation was larger, indicating that its effect was unstable and the damage to the main bud was larger. The main bud necrosis rate of the ethephon treatment group was higher, with an average of 27.6%, and the combination treatment group with boron further increased to 35.7%, and the standard deviation showed that its effect fluctuated greatly. In this study, the effects of calcium sulfate and boron fertilizer treatment at different times on the main bud necrosis rate of grape were analyzed, and the results showed that different treatment times had a significant effect on the main bud necrosis rate, and showed a clear time sequence rule Figure 11 ). Data analysis showed that the main bud necrosis rate of the May 5 treatment group was the lowest, with an average of 1.4%, and the coefficient of variation was very small, indicating that the treatment at this period not only had the best effect, but also had very high stability. In comparison, the necrosis rate of the April 26 treatment group was second (the average was 15.7%), but the data dispersion degree was higher, suggesting that the stability of the treatment effect at this period was relatively insufficient. It is worth noting that since May 25, the main bud necrosis rate showed a significant upward trend (the average increased to 23.3%), and reached a peak on June 25 (the average was as high as 45.7%), accompanied by a continuous increase in the coefficient of variation, reflecting that the effect of late treatment not only deteriorated sharply, but also the stability decreased significantly. The reason for the above time-dependent effect may be: (1) In late April, the grape bud body is in a relatively fragile stage, with low tolerance, which may lead to a relatively high necrosis rate after treatment at this period; (2) The significant increase in temperature from late May to June may indirectly increase the effective concentration of the contact agent by enhancing the volatilization rate of the calcium sulfate solution and prolonging its residence time on the bud body, thereby intensifying the risk of phytotoxicity and causing the main bud necrosis rate to rise sharply.

[0077] Table 3 Main bud necrosis, starch, germination rate, and flowering rate of different treatments

[0078]

[0079] 2.2.2 Effects of different treatments on the starch content of winter buds of main shoots

[0080] The anthrone method was used to determine the starch content in the branches. The results showed that the starch content of different treatments of sunlight rose was significantly different (Table 3), among which the starch content of the stone sulfur mixture + boron fertilizer treatment group was the highest, the data fluctuation was smaller, and it was significantly better than the water treatment, compared with the starch content before treatment, it had a significant improvement, indicating that the treatment of this group was the most conducive to starch accumulation. The performance of the stone sulfur mixture was second. The starch content of ethylene + boron fertilizer, ethylene, sodium abietate, sodium abietate + boron fertilizer was medium. The effect of water was relatively poor, among which the starch content of the treatment containing urea was low and the fluctuation was obvious, which showed that the use of high concentration urea was not conducive to the leaf nutrient return and branch nutrient accumulation under the wrong season cultivation, and from the phenotype, high concentration urea would cause the branch to dehydrate and dry up to death, so it was not recommended to use high concentration urea to promote leaf nutrient return and branch nutrient accumulation under the wrong season cultivation. Overall, the stone sulfur mixture + boron fertilizer treatment had obvious effect on improving the starch content of the branch Figure 12 ).

[0081] 2.2.3 Effect of different treatments on the winter bud germination rate of main shoots

[0082] The present application reveals significant differences in the germination process of the grape top bud by monitoring the germination dynamics of the grape top bud under different reagent combinations and concentration gradient treatments (6-21 June, every 2 days). Figure 9 , Figure 13 , Table 4). Among the reagent combination treatments, the stone sulfur mixture + boron fertilizer treatment group performed best, with the earliest germination start and the fastest germination rate, with a germination rate of 95% on June 15 (10 days after treatment) and remaining stable, which was significantly better than other treatments; in contrast, the water control group started germination late, and the final germination rate was only 88%. It is worth noting that the germination of the treatment group containing urea performed the worst, not only did it start late and progress slowly, but it also significantly reduced the final germination rate due to bud necrosis. The test results confirmed that the addition of boron fertilizer can significantly improve the germination speed, which reflects the positive regulation of calcium-boron synergy on grape germination. In the concentration gradient test, the 15 times stone sulfur mixture + 2g / L boron fertilizer treatment also performed outstandingly (Table 5), and its germination dynamics was consistent with the results of the reagent combination test: early germination start and fast peak, which was significantly different from the water control group. Figure 14 These results fully prove the synergistic effect of stone sulfur mixture and boron fertilizer, and the key influence of appropriate concentration (15 times stone sulfur mixture + 2g / L boron fertilizer) ratio on improving the germination efficiency.

[0083] Table 4 Germination rate of different treatments (unit: % / d)

[0084]

[0085] Table 5 Germination rate of different concentration treatments (unit: % / d)

[0086]

[0087] 2.2.4 Effects of different treatments on the winter bud germination rate of main shoots

[0088] The data showed that the treatment of Bordeaux mixture and its combination with boron performed the best, with the germination rate reaching 100%, which was significantly higher than that of other treatments. Ethylene treatment and water control were followed, while sodium abietate treatment showed moderate effect. It is worth noting that urea treatment performed the worst, which was due to the branch and bud necrosis caused by urea treatment. The standard deviation analysis of each treatment group showed that the stability of the Bordeaux mixture group was the highest, while the effect of the urea group was the worst. Figure 15 ).

[0089] 2.2.5 Effects of different treatments on the flower formation rate of winter buds of main shoots

[0090] The experimental results showed that there was a significant gradient difference in the effects of different treatments on the flower formation rate of grapevine. Among them, the treatment of Bordeaux mixture combined with boron fertilizer performed the best, with the flower formation rate reaching 70.65%. Sodium abietate combined with boron fertilizer (63.34%) and Bordeaux mixture alone (63.96%) were followed. Ethylene treatment, water control and sodium abietate alone showed slightly weaker effect, but ethylene combined with boron fertilizer still showed a certain synergistic trend. It is particularly worth noting that the flower formation rate of urea treatment was the worst, less than 40%. Overall, the combination of Bordeaux mixture and boron elements had a significant advantage in promoting flowering. Figure 16 ).

[0091] The present application obtains the following conclusions from the effects of different treatments on grapevine nutrient accumulation and grape sprouting and flowering under current season and off-season cultivation: (1) The combination of stone sulfur and boron fertilizer significantly promotes nutrient accumulation, and the starch content is significantly higher than that of other treatments (P<0.05). It is shown that the combination of stone sulfur and boron fertilizer promotes leaf yellowing and shedding to increase nutrient accumulation in branches. In addition, compared with single application of stone sulfur, the addition of boron fertilizer makes the dry weight ratio increase again. It further verifies that calcium and boron have a synergistic effect on the accumulation of sugar and starch. (2) The combination of stone sulfur and boron is used for the first time in the study of grape sprouting promotion, and it performs excellently, which is significantly better than other combinations in terms of sprouting speed, sprouting rate and flowering rate (P<0.05). It is shown that calcium ions can effectively promote grape sprouting, and boron elements can further improve the uniformity of sprouting by regulating nutrient allocation. (3) Based on the off-season cultivation of secondary fruits of grape, the best concentration of the combination of stone sulfur and boron fertilizer is 29% stone sulfur (15 times) + boron fertilizer (2 g / L), but attention should be paid to the temperature and time of use. The best effect is obtained in the first ten days of May in Changsha, because the sunlight rose grape bud and branch are developed well at this time, and the temperature is suitable. Too early or too late may cause bud damage. (4) Leaf removal and water treatment have poor effects. The starch content of leaf removal treatment is lower than that before treatment, and the sprouting rate and flowering rate are both lower than 75%. It is shown that early leaf removal is not conducive to nutrient accumulation, but consumes tree reserves. The indicators of water treatment are also significantly lower than those of other treatments (P<0.05), which confirms the importance of autumn leaf nutrient management. (5) Ethylene treatment performs unstably compared with the combination of stone sulfur and boron fertilizer. Although ethylene treatment can promote leaf abscission and nutrient accumulation in branches, the data fluctuates greatly, which may be related to the complex regulation of ethylene on plant endogenous hormones. Future research needs to further optimize the use method and application time. (6) The synergistic effect of boron fertilizer. Under the same treatment, the addition of boron fertilizer can improve the dry matter ratio, starch content and flowering rate. (7) Based on the off-season cultivation of secondary fruits of grape, the comparison between high-concentration urea treatment and the combination of stone sulfur and boron fertilizer shows that high-concentration urea will cause branch dehydration and dryness, and main bud necrosis, which is not conducive to the nutrient return of grape leaves and the nutrient accumulation of branches under off-season cultivation.

[0092] The present application discloses for the first time that the combination of stone sulfur and boron fertilizer has multiple positive effects on sunlight rose grape: the treatment not only significantly promotes leaf nutrient return and branch nutrient accumulation, but also effectively improves the sprouting speed and uniformity. It is particularly innovative that the present application successfully applies the treatment scheme containing stone sulfur to the production practice of secondary fruits of sunlight rose grape under off-season cultivation for the first time. The results show that the combination of stone sulfur and boron fertilizer can be used as an innovative technology to improve the sprouting quality of grape under conventional cultivation, and more effectively promote nutrient accumulation and flower bud differentiation under off-season cultivation.

Claims

1. The application of a combination agent for promoting grape bud break in promoting grape bud break, characterized in that: The combination agent includes lime-sulfur mixture and boron fertilizer.

2. The application according to claim 1, characterized in that: In the combined agent, the lime-sulfur mixture is 10-20 times the concentration of 29% lime-sulfur mixture, and the boron fertilizer is 1g / L-4g / L.

3. The application according to claim 2, characterized in that: In the combined agent, the lime-sulfur mixture is 15 times the concentration of 29% lime-sulfur mixture, and the boron fertilizer is 2 g / L.

4. A method for promoting grape bud break using a combination of agents that promote grape bud break, characterized in that, Spray the entire grapevine with the aforementioned combination agent 15 days before leaf fall under the regular season cultivation model or more than 30 days before bud break under the off-season cultivation model based on secondary fruit. The combined agent includes lime-sulfur mixture and boron fertilizer.

5. The method according to claim 4, characterized in that: In the combined agent, the lime-sulfur mixture is 10-20 times the concentration of 29% lime-sulfur mixture, and the boron fertilizer is 1g / L-4g / L.

6. The method according to claim 5, characterized in that: In the combined agent, the lime-sulfur mixture is 15 times the concentration of 29% lime-sulfur mixture, and the boron fertilizer is 2 g / L.

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