Preparation of biocontrol bacterium compound agent and application of biocontrol bacterium compound agent to grape coloring

Abstract

The invention provides preparation of a biocontrol bacterium compound agent and application of the biocontrol bacterium compound agent to grape coloring, and the preparation method comprises the following steps: controlling the pH value and culture temperature of bacillus velezensis for culture, controlling the rotating speed, the ventilation capacity and the pressure in the logarithmic phase for culture, and adjusting the rotating speed, the ventilation capacity and the pressure in the stable phase for continuous fermentation culture; the cultured bacillus velezensis is mixed with COR coronatine (0.1%-0.5%), 5-aminolevulinic acid (0.2%-0.4%), 1-aminocyclopropanecarboxylic acid (0.15%-0.3%) and the bacillus velezensis (1%-3%), and the biocontrol bacterium compound agent is obtained. The biocontrol bacterium compound agent disclosed by the invention is high in concentration, small in usage amount, low in cost and small in influence on the environment; meanwhile, the biocontrol bacterium compound agent can effectively promote color change of the grapes and improve the commodity value of the grapes; and the biocontrol bacterium compound agent does not influence the activity of the biocontrol bacterium after being stored for a long time, so that the color changing period of the grapes is shortened, and the quality and yield of the grapes are improved.

Description

Technical Field

[0001] The invention relates to the technical field of grape color change, and in particular to the preparation of a biocontrol bacteria compound and its application to grape color change. Background Art

[0002] The quality of grape fruit is mainly affected by many factors, among which the color of the fruit is one of the important indicators for judging the quality of the fruit. Traditionally, the color of grape fruit is mainly regulated by endogenous hormones, such as jasmonic acid, ethylene, abscisic acid, etc. These endogenous hormones affect the color formation of the fruit by regulating the synthesis and transport of pigments in the fruit. However, due to the influence of factors such as the growth environment, climate and cultivation management, the color change process of grape fruit may be restricted to a certain extent, resulting in uneven fruit color or slow fruit color change, affecting the appearance and market competitiveness of the fruit.

[0003] In order to solve the problems existing in the color change process of grape berries, many researchers are committed to finding new control methods and strategies to increase the color change speed and uniformity of grape berries and improve the appearance and quality of the berries. In recent years, biostimulants have attracted much attention as a new type of fruit color change regulator. Compared with traditional chemical color adjusters, biostimulants have the advantages of being environmentally friendly, non-toxic, and harmless to human health, so they have been widely studied and applied.

[0004] At present, there is still a lack of a biostimulant compound that can effectively promote the color change of grape fruit on the market. Plant biostimulants stimulate the natural process of plants and have nothing to do with nutritional components. Studies have found that spraying seaweed fertilizer SUNRED improves the fruit skin color index CIRG of "Red Globe" fruit, increases the content of anthocyanins, and promotes coloring; spraying humic acid "Palm" and seaweed extract "Sea Love Plus" increases the anthocyanin content of "Summer Black" grapes by 65.6% and 36.2% respectively during the mature period.

[0005] Although existing biostimulant products can promote fruit color change to a certain extent, their effects are limited, and there are problems such as complex use and unstable effects. Therefore, it is of great scientific research and application value to find a new type of biostimulant compound that can effectively promote the color change process of grape fruit and improve the color uniformity and quality of the fruit.

[0006] Therefore, the present invention provides a preparation of a biocontrol bacteria compound and its application to grape color change. The biostimulant compound of the present invention can effectively promote the color change process of grape fruit, improve the appearance and quality of the fruit, and provide new technical support and solutions for the development of the grape industry. Summary of the invention

[0007] The invention aims to provide a preparation method of a biocontrol bacteria compound and its application to grape color change, so as to promote the color change process of grape fruit and improve the appearance and quality of the fruit.

[0008] The purpose of the present invention is achieved through the following technical solutions:

[0009] A method for preparing a biocontrol agent, wherein the biocontrol agent is prepared by mixing a biostimulant with a microbial strain, wherein the microbial strain is Bacillus velez; the biostimulant comprises: coronatine COR, 5-aminolevulinic acid (5-ALA), and 1-aminocyclopropanecarboxylic acid;

[0010] The preparation method of the biocontrol bacteria compound comprises the following steps:

[0011] (1) controlling the pH value and the culture temperature of Bacillus velezii, controlling the rotation speed, the ventilation volume and the pressure during the logarithmic growth period, and adjusting the rotation speed, the ventilation volume and the pressure during the stable period to continue the fermentation culture;

[0012] (2) The Bacillus Velez subtilis fermented and cultured in step (1) is mixed with the COR coronatine, 5-aminolevulinic acid (5-ALA), and 1-aminocyclopropanecarboxylic acid to obtain the biocontrol bacteria compound.

[0013] Preferably, the mass percentage of COR coronatine in the biocontrol bacteria compound is 0.1% to 0.5%; the mass percentage of 5-aminolevulinic acid (5-ALA) is 0.2% to 0.4%; the mass percentage of 1-aminocyclopropanecarboxylic acid is 0.15% to 0.3%; and the mass percentage of Bacillus velez is 1% to 3%.

[0014] Preferably, in step (1), the pH value of the Bacillus Velez subtilis is controlled to be between 7 and 9, and the culture temperature is 30 to 34°C.

[0015] Preferably, in step (1), the rotation speed during the logarithmic growth phase is controlled to be 75-85 rpm, the ventilation volume is 75-85 ml / min, and the pressure is 0.06-0.07 MPa for culturing for 10-16 hours.

[0016] Preferably, in step (1), during the stable period, the rotation speed is adjusted to 115-130 rpm, the ventilation volume is adjusted to 115-130 ml / min, and the pressure is adjusted to 0.08-0.09 MPa to continue the fermentation culture for 48-56 hours.

[0017] The present application also claims protection for a biocontrol bacteria compound prepared by the above-mentioned biocontrol bacteria compound preparation method.

[0018] The present application also claims to protect the use of the above-mentioned biocontrol bacteria compound in grape color change.

[0019] In the above technical solution, COR coronatine can induce plant gene expression, activate biosynthetic pathways of anthocyanins and anthocyanins, increase the accumulation of high levels of anthocyanins in the peel and pulp, and at the same time increase the accumulation of proteins, amino acids, sugars and other substances in the plant by increasing the photosynthetic rate of crops. In simple terms, it can change color, increase sugar and reduce acid.

[0020] In the above technical scheme, 5-aminolevulinic acid (5-ALA) is closely related to the synthesis and accumulation of anthocyanins in fruits and vegetables. 5-ALA can effectively promote the expression of related genes, thereby promoting the early synthesis and accumulation of anthocyanins in the peel, promoting fruit coloring; accelerating the conversion of substances accumulated by photosynthesis into soluble sugars, increasing sugar accumulation, and improving fruit quality.

[0021] In the above technical solution, 1-aminocyclopropanecarboxylic acid improves the efficiency of grape color change and accelerates the ripening process of the fruit by promoting the synthesis of ethylene, regulating the expression of genes related to pigment synthesis, accelerating the degradation of chlorophyll, and promoting sugar accumulation. However, in actual applications, 1-aminocyclopropanecarboxylic acid needs to be used with caution, and excessive use may have an adverse effect on fruit quality and plant growth.

[0022] Due to the application of the above technical solution, the present invention has the following beneficial effects compared with the prior art:

[0023] The biocontrol bacteria compound of the present invention has high concentration, small usage amount, low cost and little impact on the environment; at the same time, the biocontrol bacteria compound can effectively promote the color change of grapes and improve the commercial value of grapes; and the biocontrol bacteria compound of the present invention does not affect the activity of the biocontrol bacteria even after long-term storage, shortens the color change period of grapes and improves the quality and yield of grapes. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] In order to more clearly illustrate the specific implementation methods of the present invention or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, some of the drawings described below are some implementation methods of the present invention. For ordinary technicians in this field, other drawings can be made based on these drawings without paying any creative work.

[0025] Figure 1 It is a physical picture of formula 3 and the control group in Example 1 of the present invention. DETAILED DESCRIPTION

[0026] In order to have a clearer understanding of the technical features, purposes and effects of the present invention, a specific implementation scheme is now described in detail.

[0027] The present invention is further described below in conjunction with the embodiments, but the present invention is not limited to the following embodiments. The implementation conditions used in the embodiments can be further adjusted according to the different requirements of specific use, and the implementation conditions not indicated are conventional conditions in the industry. The technical features involved in each embodiment of the present invention can be combined with each other as long as they do not conflict with each other.

[0028] Example 1

[0029] See attached Figure 1 This embodiment provides a method for preparing a biocontrol agent, wherein the biocontrol agent is prepared by mixing a biostimulant with a microbial strain, wherein the microbial strain is Bacillus velez; the biostimulant includes: coronatine COR, 5-aminolevulinic acid (5-ALA), and 1-aminocyclopropanecarboxylic acid;

[0030] The preparation method of the biocontrol bacteria compound comprises the following steps:

[0031] (1) Bacillus Velezii was cultured at a pH value of 7.2 and a culture temperature of 32°C. The rotation speed was controlled at 80 rpm, the ventilation volume was 80 ml / min, and the pressure was 0.07 MPa during the logarithmic growth phase for 12 h. The rotation speed was adjusted to 120 rpm, the ventilation volume was 120 ml / min, and the pressure was 0.08 MPa during the stable phase and the fermentation culture was continued for 48 h.

[0032] (2) mixing the Bacillus velez fermented and cultured in step (1) with the COR coronatine, 5-aminolevulinic acid (5-ALA), and 1-aminocyclopropanecarboxylic acid to obtain the biocontrol bacteria composite agent;

[0033] Specifically, the compounding ratio of the biocontrol bacteria compound is shown in Table 1.

[0034] Table 1

[0035] Components Recipe 1 Recipe 2 Recipe 3 Bacillus Velez 1％ 2％ 3％ Coronatine 0.1％ 0.5％ 0.3％ 5-Aminolevulinic acid (5-ALA) 0.2％ 0.3％ 0.4％ 1-Aminocyclopropanecarboxylic acid 0.15％ 0.3％ 0.2％

[0036] The above-mentioned biocontrol agent was used during the grape color change period, and the CK group without the biocontrol agent was used as a blank control group for testing. The test results are shown in Table 2.

[0037] Table 2

[0038] Components Recipe 1 Recipe 2 Recipe 3 CK Single string coloring ratio 99.8 100 100 90 Total soluble sugar per string 22.98 23.25 24.8 18.6 Single bunch fruit weight g 400 450 480 320 Single fruit weight g 8.5 9.8 9.1 6.5 Number of fruits (bunch) 16 16 17 14

[0039] As can be seen from Table 2 above, the biocontrol bacteria compound agents of different biostimulants show that when applied during the grape color change period, the coloring ratio of a single bunch can be increased by 10.89% to 11.11%, the total soluble sugar of a single bunch can be increased by 23.55 to 33.33%, the fruit weight of a single bunch can be increased by 25% to 50%, the single fruit weight can be increased by 30.77 to 50.77%, and the number of fruits can be increased by 14.29% to 21.43% compared with the control group without application. Comprehensively compared, formula 3 (3% Bacillus velez + 0.3% coronatine + 0.4% 5-aminolevulinic acid + 0.2% 1-aminocyclopropanecarboxylic acid) has the best effect and can be used as the best biocontrol bacteria compound agent for application during the grape color change period.

[0040] Stability test of the above biocontrol compound:

[0041] The above-mentioned biocontrol bacteria compound was stored at 4°C, 28°C, 37°C, and 40°C, and the color and odor changes were observed, and the number of viable bacteria was detected. The color change is shown in Table 3, the odor change is shown in Table 4, and the number of viable bacteria is shown in Table 5.

[0042] Table 3

[0043]

[0044] Table 4

[0045]

[0046] Table 5

[0047]

[0048] As can be seen from the above table, the present invention has found through research that the high-concentration biocontrol bacteria compound agent composed of COR coronatine (0.1% to 0.5%), 5-aminolevulinic acid (0.2% to 0.4%), 1-aminocyclopropanecarboxylic acid (0.15% to 0.3%) and Bacillus velez (1% to 3%) is beneficial to the increase of single-bunch coloring ratio by 10.89% to 11.11%, the increase of single-bunch total soluble sugar by 23.55 to 33.33%, the increase of single-bunch fruit weight by 25% to 50%, the increase of single fruit weight by 30.77 to 50.77%, and the increase of fruit number by 14.29% to 21.43% when applied during the grape color change period. Formula 3 has the best comprehensive benefit. At the same time, long-term storage does not affect the activity of the biocontrol bacteria, shortens the grape color change period and improves the quality and yield of the grape.

[0049] In summary, the biocontrol bacteria compound of the present invention has high concentration, small usage amount, low cost, and little impact on the environment; at the same time, the biocontrol bacteria compound can effectively promote the color change of grapes and improve the commercial value of grapes; and the biocontrol bacteria compound of the present invention does not affect the activity of the biocontrol bacteria during long-term storage, shortens the color change period of grapes and improves the quality and yield of grapes.

[0050] The above-mentioned embodiment only expresses one implementation mode of the present invention, and its description is relatively specific and detailed, but it cannot be understood as limiting the scope of the invention patent. It should be pointed out that for ordinary technicians in this field, several modifications and improvements can be made without departing from the concept of the present invention, which all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be based on the attached claims.

Claims

1. A method for preparing a biocontrol bacteria compound, characterized in that: The biocontrol bacteria compound is prepared by mixing a biostimulant with a microbial strain, wherein the microbial strain is Bacillus velez; the biostimulant includes: COR coronatine, 5-aminolevulinic acid, and 1-aminocyclopropanecarboxylic acid; The preparation method of the biocontrol bacteria compound comprises the following steps: (1) controlling the pH value and the culture temperature of Bacillus velezii, controlling the rotation speed, the ventilation volume and the pressure during the logarithmic growth period, and adjusting the rotation speed, the ventilation volume and the pressure during the stable period to continue the fermentation culture; (2) Mixing the Bacillus Velez subtilis fermented in step (1) with the COR coronatine, 5-aminolevulinic acid, and 1-aminocyclopropanecarboxylic acid to obtain the biocontrol bacteria compound.

2. The method for preparing a biocontrol agent according to claim 1, characterized in that: The mass percentage of COR coronatine in the biocontrol bacteria compound is 0.1% to 0.5%; the mass percentage of 5-aminolevulinic acid is 0.2% to 0.4%; the mass percentage of 1-aminocyclopropanecarboxylic acid is 0.15% to 0.3%; and the mass percentage of Bacillus velez is 1% to 3%.

3. The method for preparing a biocontrol agent according to claim 1, characterized in that: In step (1), the pH value of the Velez subtilis is controlled to be between 7 and 9, and the culture temperature is 30 to 34°C.

4. The method for preparing a biocontrol agent according to claim 1, characterized in that: In step (1), during the logarithmic growth phase, the rotation speed is controlled to be 75-85 rpm, the ventilation volume is 75-85 ml / min, and the pressure is 0.06-0.07 MPa for 10-16 hours.

5. The method for preparing a biocontrol agent according to claim 1, characterized in that: In step (1), during the stable period, the rotation speed is adjusted to 115-130 rpm, the ventilation volume is adjusted to 115-130 ml / min, and the pressure is adjusted to 0.08-0.09 MPa to continue the fermentation culture for 48-56 hours.

6. The biocontrol bacteria compound prepared by the method for preparing the biocontrol bacteria compound according to any one of claims 1 to 5.

7. Use of the biocontrol bacteria compound prepared by the preparation method of the biocontrol bacteria compound according to any one of claims 1 to 5 or the biocontrol bacteria compound according to claim 6 in grape color change.

Images