Composite probiotics with cucumber growth-promoting and yield-increasing effects and application thereof

Abstract

The invention relates to compound probiotics with cucumber growth-promoting and yield-increasing effects and application of the compound probiotics. The compound probiotics with the cucumber growth promoting and yield increasing effects are prepared from a bacillus coagulans BC66 strain with the preservation number of CGMCC (China General Microbiological Culture Collection Center) 21879, a lactobacillus plantarum Lp90 strain with the preservation number of CGMCC No.10453, a pediococcus acidilactici PA153 strain with the preservation number of CCTCC NO: M 2023230, and a lactobacillus acidophilus LA71 strain with the preservation number of CGMCC 21765, and the compound probiotics with the cucumber growth promoting and yield increasing effects are prepared from the following raw materials: the bacillus coagulans BC66 strain, the lactobacillus plantarum Lp90 strain, the pediococcus acidilactici PA153 strain and the lactobacillus acidophilus LA71 strain. The four strains have a remarkable synergistic effect in the aspects of growth promotion and yield increase of cucumbers.

Description

Technical Field

[0001] The invention belongs to the technical field of microorganisms and relates to a composite probiotic having the effect of promoting growth and increasing yield of cucumber and an application thereof. Background Art

[0002] Cucumber is an important vegetable crop with high yield, early fruiting, quick effect and long supply period. However, during the cultivation and growth of cucumber, it is easily affected by various factors such as environment, soil and fertilizer, resulting in unstable yield and quality of cucumber. Although traditional chemical fertilizers can increase yield to a certain extent, long-term and large-scale use will have a negative impact on the soil and ecological environment.

[0003] Probiotics are living microorganisms that can not only increase soil microorganisms, inhibit the growth of soil pathogens, loosen the soil, and enhance crop photosynthesis, but also supplement the organic nutrients of crops by their metabolites, thereby enhancing plant growth, promoting root growth, increasing yields, and improving crop quality. Therefore, it is of great significance to develop a probiotic product that can effectively promote growth and increase cucumber yields. Summary of the invention

[0004] In view of the deficiencies of the prior art, the object of the present invention is to provide a composite probiotic having the effect of promoting the growth and increasing the yield of cucumbers and its application.

[0005] In order to achieve the purpose of the invention, the present invention adopts the following technical solutions:

[0006] In a first aspect, the present invention provides a composite probiotic having the effect of promoting growth and increasing yield of cucumbers, wherein the composite probiotic having the effect of promoting growth and increasing yield of cucumbers is composed of Bacillus coagulans with a preservation number of CGMCC 21879. Bacillus coagulating BC66 strain, Lactobacillus plantarum with a deposit number of CGMCC No.10453 Lactobacillus plants Lp90 strain, Pediococcus acidilactici with a deposit number of CCTCC NO: M 2023230 Pediococcus lactic acid PA153 strain and Lactobacillus acidophilus with the deposit number CGMCC 21765 Lactobacillus acidophilus LA71 strain composition.

[0007] The present invention has developed a new probiotic compounding method and a new strategy for promoting the growth and increasing the yield of cucumbers, namely, Bacillus coagulans BC66 strain, Lactobacillus plantarum Lactobacillus plantarum Lp90 strain, Pediococcus acidilactici Pediococcus acidilactici PA153 strain and Lactobacillus acidophilus Lactobacillus acidophilus The LA71 strain was compounded and it was found that there was a potential interaction between the four strains, which could cooperate with each other and synergize. When the amount of bacteria was the same, compared with the intervention method without any bacteria, the effect of the compound of the four bacteria in promoting the growth and yield of cucumbers was significantly improved, which was specifically manifested in increasing the height of cucumber plants, stem thickness and yield. Therefore, the use of this composite probiotic in the field of cucumber cultivation has a good prospect.

[0008] The preparation method of the composite probiotics can be prepared by conventional technical methods in the art, and can be exemplified by: activating the four strains, inoculating them into a culture medium for cultivation, obtaining a culture solution; centrifuging the culture solution, resuspending the bacteria to obtain a bacterial suspension; and mixing the four bacterial suspensions according to the ratio of the number of live bacteria. Alternatively, a protective agent is further added for freeze drying to obtain a freeze-dried bacterial powder product.

[0009] Preferably, the culture medium comprises MRS medium.

[0010] Preferably, the MRS medium comprises, by concentration, the following: peptone 8-12 g / L, beef extract 8-12 g / L, glucose 15-25 g / L, sodium acetate 1-3 g / L, yeast powder 3-7 g / L, diammonium hydrogen citrate 1-3 g / L, K2PO4·3H2O 2-3 g / L, MgSO4·7H2O 0.05-0.2 g / L, MnSO4 0.01-0.1 g / L, Tween 80 0.5-2 mL / L, and cysteine ​​hydrochloride 0.1-1 g / L.

[0011] Preferably, the ratio of the viable counts of the BC66 strain, the Lp90 strain, the PA153 strain and the LA71 strain is (1-10):(1-10):(1-10):(1-10);

[0012] Among them, the above-mentioned "1-10" can be, for example, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, etc. Other specific point values ​​within this numerical range can be selected, and they will not be elaborated here one by one.

[0013] Based on the potential interaction between the four strains, the present invention also found that when the four strains are compounded and used in the above-mentioned specific ratio of viable bacteria count, the effect on promoting the growth and increasing the yield of cucumber is more significant.

[0014] In a second aspect, the present invention provides a probiotic agent having the effect of promoting growth and increasing yield of cucumbers, wherein the strain in the probiotic agent includes the composite probiotic described in the first aspect.

[0015] Preferably, the live bacterial content of the BC66 strain, Lp90 strain, PA153 strain and LA71 strain in the probiotic agent is not less than 1×10 8 CFU / mL or 1×10 8 CFU / g, for example 1×10 8 CFU / mL (CFU / g), 2×10 8 CFU / mL (CFU / g), 5×10 8 CFU / mL (CFU / g), 8×10 8 CFU / mL (CFU / g), 1×10 9 CFU / mL (CFU / g), 5×10 9 CFU / mL (CFU / g), 1×10 10 CFU / mL (CFU / g), etc. Other specific point values ​​within this numerical range can be selected and will not be described here one by one.

[0016] Preferably, the dosage form of the probiotic includes solution, lyophilized powder, capsule, tablet or granule.

[0017] Preferably, the probiotic further comprises a lyophilization protectant.

[0018] Preferably, the lyoprotectant comprises any one of skim milk, sucrose, lactose, trehalose, dextran, gelatin, dextrin, gum arabic, sodium alginate, polyvinyl pyrrolidone, sorbitol or xylitol, or a combination of at least two thereof.

[0019] In a third aspect, the present invention provides a use of the composite probiotics according to the first aspect or the probiotic agent according to the second aspect in cucumber planting.

[0020] In a fourth aspect, the present invention provides a cucumber growth-promoting and yield-increasing fertilizer, wherein the cucumber growth-promoting and yield-increasing fertilizer comprises the composite probiotics described in the first aspect or the probiotic agent described in the second aspect.

[0021] Preferably, the total viable count of probiotics in the fertilizer is not less than 1×10 6 CFU / mL or 1×10 6 CFU / g, for example, can be 1×10 6 CFU / mL (CFU / g), 2×10 6 CFU / mL (CFU / g), 5×10 6 CFU / mL (CFU / g), 8×10 6 CFU / mL (CFU / g), 1×10 7 CFU / mL (CFU / g), 5×10 7CFU / mL (CFU / g), 1×10 8 CFU / mL (CFU / g), etc. Other specific point values ​​within this numerical range can be selected and will not be described here one by one.

[0022] Compared with the prior art, the present invention has the following beneficial effects:

[0023] The present invention has developed a new probiotic compounding method and a new strategy for promoting the growth and increasing the yield of cucumbers, namely, Bacillus coagulans BC66 strain, Lactobacillus plantarum Lactobacillus plantarum Lp90 strain, Pediococcus acidilactici Pediococcus acidilactici PA153 strain and Lactobacillus acidophilus Lactobacillus acidophilus The LA71 strain was compounded and it was found that there was a potential interaction between the four strains, which could cooperate with each other and synergize. When the amount of bacteria was the same, compared with the intervention method without any bacteria, the effect of the compound of the four bacteria in promoting the growth and yield of cucumbers was significantly improved, which was specifically manifested in increasing the height of cucumber plants, stem thickness and yield. Therefore, the use of this composite probiotic in the field of cucumber cultivation has a good prospect.

[0024] The BC66 strain involved in the present invention is classified as Bacillus coagulans Bacillus coagulans The depository is the General Microbiology Center of China Microorganism Culture Collection Administration, the deposit number is CGMCC 21879, the deposit date is March 8, 2021, and the deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0025] The Lp90 strain involved in the present invention is classified and named as Lactobacillus plantarum Lactobacillus plantarum The depository is the General Microbiology Center of China Microorganism Culture Collection Administration, the deposit number is CGMCC No.10453, the deposit date is January 27, 2015, and the deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0026] The PA153 strain involved in the present invention is classified and named as Pediococcus acidilactici PA153 Pediococcus lactic acid PA153, the depository is China Center for Type Culture Collection, the deposit number is CCTCC NO: M2023230, the deposit date is March 2, 2023, and the deposit address is Wuhan University, Wuhan, China.

[0027] The LA71 strain involved in the present invention is classified and named as Lactobacillus acidophilus Lactobacillus acidophilusThe depository is the General Microbiology Center of China Microorganism Culture Collection Administration, the deposit number is CGMCC 21765, the deposit date is January 29, 2021, and the deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 It is a statistical chart of the results of two measurements of the height of cucumber seedlings in each group;

[0029] Figure 2 It is a statistical chart of the results of two measurements of the stem diameter of cucumber seedlings in each group;

[0030] Figure 3 It is a statistical chart of the measurement results of cucumber fruit yield in each group. DETAILED DESCRIPTION

[0031] In order to further explain the technical means and effects adopted by the present invention, the technical solution of the present invention is further described below in conjunction with the preferred embodiments of the present invention, but the present invention is not limited to the scope of the embodiments.

[0032] The bacterial species involved in the following embodiments are:

[0033] ① The BC66 strain was named Bacillus coagulans Bacillus coagulans , the deposit number is CGMCC21879;

[0034] ② The classification of Lp90 strains is named Lactobacillus plantarum Lactobacillus plantarum , the deposit number is CGMCC No.10453;

[0035] ③ The PA153 strain was named as Pediococcus acidilactici PA153 Pediococcus acidilactici PA153, deposit number is CCTCC NO: M 2023230;

[0036] ④ The LA71 strain was named Lactobacillus acidophilus Lactobacillus acidophilus , the deposit number is CGMCC 21765;

[0037] ⑤ ATCC 7050 strain is Bacillus coagulans ATCC 7050 strain.

[0038] The culture medium formula involved in the following examples is as follows:

[0039] Peptone 10g / L, beef extract 10g / L, glucose 20g / L, sodium acetate 2g / L, yeast powder 5g / L, diammonium hydrogen citrate 2g / L, K2PO4·3H2O 2.6g / L, MgSO4·7H2O 0.1g / L, MnSO4 0.05g / L, Tween 80 1mL / L, cysteine ​​salt 0.5g / L.

[0040] The following embodiment involves a method for preparing a bacterial suspension: inoculating the desired strain into a liquid culture medium, culturing at 37°C for 24 h for activation, and continuously activating twice to obtain an activated solution; inoculating the activated solution into a liquid culture medium at an inoculum amount of 5% (v / v), culturing at 37°C for 24 h to obtain a bacterial solution; centrifuging the bacterial solution at 4°C and 5000 rpm for 10 min, filtering to obtain bacterial bodies, and resuspending the bacterial bodies in a PBS solution to obtain the obtained product.

[0041] Example 1

[0042] The present embodiment provides a composite probiotic, which is composed of a BC66 bacterial suspension, an Lp90 bacterial suspension, a PA153 bacterial suspension, and a LA71 bacterial suspension in a ratio of 1:1:1:1, wherein the concentration of the live bacteria count of BC66 in the composite probiotic is 6×10 8 CFU / mL.

[0043] Example 2

[0044] The present embodiment provides a composite probiotic, which is composed of a BC66 bacterial suspension, an Lp90 bacterial suspension, a PA153 bacterial suspension, and a LA71 bacterial suspension in a ratio of 2:3:5:2, wherein the concentration of the live bacteria count of BC66 in the composite probiotic is 4×10 8 CFU / mL.

[0045] Example 3

[0046] The present embodiment provides a composite probiotic, which is composed of a BC66 bacterial suspension, an Lp90 bacterial suspension, a PA153 bacterial suspension, and a LA71 bacterial suspension in a ratio of 7:2:6:9 in terms of viable bacterial count, wherein the viable bacterial count concentration of BC66 in the composite probiotic is 7×10 8 CFU / mL.

[0047] Comparative Example 1

[0048] This comparative example provides a composite probiotic, which differs from Example 1 only in that the BC66 bacterial suspension is replaced by an ATCC 7050 bacterial suspension with an equal number of viable cells.

[0049] Comparative Example 2

[0050] This comparative example provides a composite probiotic, which differs from Example 1 only in that it does not contain the BC66 strain, and its reduced amount is distributed to the Lp90 strain, the PA153 strain and the LA71 strain according to the ratio of the number of live bacteria, and the total number of live bacteria in the composite probiotic is kept equal to that in Example 1.

[0051] Comparative Example 3

[0052] This comparative example provides a composite probiotic, which differs from Example 1 only in that it does not contain the Lp90 strain, and its reduced amount is distributed to the BC66 strain, the PA153 strain and the LA71 strain according to the ratio of the number of live bacteria, and the total number of live bacteria in the composite probiotic is kept equal to that in Example 1.

[0053] Comparative Example 4

[0054] This comparative example provides a composite probiotic, which differs from Example 1 only in that it does not contain the PA153 strain, and its reduced amount is distributed to the BC66 strain, the Lp90 strain and the LA71 strain according to the ratio of the number of live bacteria, and the total number of live bacteria in the composite probiotic is kept equal to that in Example 1.

[0055] Comparative Example 5

[0056] This comparative example provides a composite probiotic, which differs from Example 1 only in that it does not contain the LA71 strain, and its reduced amount is distributed to the BC66 strain, the Lp90 strain and the PA153 strain according to the ratio of the number of live bacteria, and the total number of live bacteria in the composite probiotic is kept equal to that in Example 1.

[0057] Test Case

[0058] This test example tests the growth-promoting and yield-increasing effects of the composite probiotics prepared in Examples 1-3 and Comparative Examples 1-5 on cucumbers.

[0059] (1) Sample processing: The composite probiotics prepared in Examples 1-3 and Comparative Examples 1-5 were mixed with water, diluted 300 times, stirred and mixed, and then used.

[0060] (2) Plant samples: Cucumber seedlings at 15 days into the seedling stage, plant height: 35±1.0 cm, stem diameter: 0.7±0.05 cm.

[0061] (3) Planting conditions: temperature controlled at 24-28℃, humidity 70-90%, sunshine: 10 h.

[0062] (4) Application method: The first root irrigation of cucumber seedlings (seedling stage 15 days) was carried out with the composite probiotics of each group diluted and mixed in step (1), and the second root irrigation was carried out after an interval of 7 days. The third root irrigation was carried out after an interval of 7 days after the second root irrigation. The amount of the composite probiotics diluted solution for each root irrigation was 2.5 kg / mu. At the same time, a blank control group was set up, and the root irrigation was carried out three times at the same time with the same amount of water. Each treatment group treated one ridge and two rows of cucumber seedlings, and the plants of each group were cultured under the same environmental conditions.

[0063] (5) The plant height and stem diameter of the cucumber seedlings were measured and counted twice, 9 days after the first root irrigation and 16 days after the first root irrigation. The yield of each group was counted after the fruits matured. The results were presented as average values, where plant height, stem diameter and yield were as follows: Figure 1 , Figure 2 and Figure 3 shown.

[0064] From the statistical results in the figure, we can see that:

[0065] (1) It can be seen from Examples 1 to 3 that the present invention uses four strains, namely BC66, Lp90, PA153 and LA71, for compound use, which can effectively promote the growth of cucumber seedlings. The plant height and stem thickness are significantly better than those of the blank control group, and the fruit yield is greatly improved.

[0066] (2) By comparing Example 1 with Comparative Examples 1-5, it can be seen that when the commercially available Bacillus coagulans is used to replace the BC66 strain in the present invention in Comparative Example 1, the effect of the composite probiotic solution prepared therefrom on promoting the growth and increasing the yield of cucumber plants after application is lower than that in Example 1; and when Comparative Examples 2-5 do not contain any of the four strains of BC66, Lp90, PA153 and LA71, the plant height and stem thickness of the cucumber seedlings after application of the composite probiotic solution prepared therefrom are not as good as those in Example 1, and the fruit yield is also not as good as that in Example 1. It can be seen that the combined use of the four strains of BC66, Lp90, PA153 and LA71 in the present invention can effectively enhance the growth ability of cucumber plants, promote root growth and seedling growth, and increase yield.

[0067] In summary, the present invention has developed a new probiotic compounding method and a new strategy for promoting the growth and increasing the yield of cucumbers, namely, Bacillus coagulans BC66 strain, Lactobacillus plantarum Lactobacillus plants Lp90 strain, Pediococcus acidilactici Pediococcus acidilactici PA153 strain and Lactobacillus acidophilus Lactobacillus acidophilusThe LA71 strain was compounded and it was found that there was a potential interaction between the four strains, which could cooperate with each other and synergize. When the amount of bacteria was the same, compared with the intervention method without any bacteria, the effect of the compound of the four bacteria in promoting the growth and yield of cucumbers was significantly improved, which was specifically manifested in increasing the height of cucumber plants, stem thickness and yield. Therefore, the use of this composite probiotic in the field of cucumber cultivation has a good prospect.

[0068] The applicant declares that the present invention illustrates the technical solution of the present invention through the above-mentioned embodiments, but the present invention is not limited to the above-mentioned embodiments, that is, it does not mean that the present invention must rely on the above-mentioned embodiments to be implemented. Those skilled in the art should understand that any improvement of the present invention, equivalent replacement of various raw materials of the product of the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

[0069] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, a variety of simple modifications can be made to the technical solution of the present invention, and these simple modifications all belong to the protection scope of the present invention.

[0070] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations.

Claims

1. A composite probiotic having the effect of promoting growth and increasing yield of cucumber, characterized in that: The composite probiotics with cucumber growth-promoting and yield-increasing effects are prepared from Bacillus coagulans with a preservation number of CGMCC 21879. Bacillus coagulans BC66 strain, Lactobacillus plantarum with a deposit number of CGMCC No.10453 Lactobacillus plantarum Lp90 strain, Pediococcus acidilactici with a deposit number of CCTCC NO: M 2023230 Pediococcus acidilactici PA153 strain and Lactobacillus acidophilus with the deposit number CGMCC 21765 Lactobacillus acidophilus LA71 strain composition.

2. The composite probiotic having the effect of promoting growth and increasing yield of cucumber according to claim 1, characterized in that: The ratio of the viable bacterial counts of the BC66 strain, the Lp90 strain, the PA153 strain and the LA71 strain is (1-10):(1-10):(1-10):(1-10).

3. A probiotic agent having the effect of promoting growth and increasing yield of cucumber, characterized in that: The strains in the probiotics include the composite probiotics according to claim 1 or 2.

4. The probiotic according to claim 3, characterized in that The live bacterial content of the BC66 strain, the Lp90 strain, the PA153 strain and the LA71 strain in the probiotic agent is not less than 1×10 8 CFU / mL or 1×10 8 CFU / g.

5. The probiotic according to claim 3, characterized in that The dosage form of the probiotics includes solution, lyophilized powder, capsule, tablet or granule.

6. The probiotic according to claim 3, characterized in that The probiotics also include a lyoprotectant.

7. The probiotic according to claim 6, characterized in that The lyoprotectant includes any one of skim milk, sucrose, lactose, trehalose, dextran, gelatin, dextrin, gum arabic, sodium alginate, polyvinyl pyrrolidone, sorbitol or xylitol, or a combination of at least two thereof.

8. Use of the composite probiotic according to claim 1 or 2 or the probiotic agent according to any one of claims 3 to 7 in cucumber planting.

9. A cucumber growth-promoting and yield-increasing fertilizer, characterized in that: The cucumber growth-promoting and yield-increasing fertilizer comprises the composite probiotics described in claim 1 or 2 or the probiotic agent described in any one of claims 3-7.

10. The cucumber growth-promoting and yield-increasing fertilizer according to claim 9, characterized in that: The total viable count of probiotics in the fertilizer is not less than 1×10 6 CFU / mL or 1×10 6 CFU / g.

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