A microbial compound bacterial agent and its preparation method and application
By developing a microbial complex agent containing Bassia zoluma and Serratia sarcoplasma, the problems of continuous cropping and soil salinization in tomato planting were solved, and the soil environment improvement and tomato growth promotion were achieved.
Patent Information
- Application Number
- CN202411687347.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2024-11-25
- Publication Date
- 2025-05-23
- Estimated Expiration
- 2044-11-25
AI Technical Summary
There are continuous cropping obstacles and soil salinization problems in tomato planting, resulting in soil environmental deterioration and plant growth limitation.
A microbial complex agent, including Sporosarcina saromensis ZIP-102 and Serratia marcescens GRSB-147, was developed, prepared by mixing its fermentation broth and applied to the soil to improve the soil environment.
This microbial compound bacteria agent can effectively improve the soil environment, increase the height, stem thickness and fresh weight of tomato plants, reduce the use of chemical fertilizers, and have the development prospects for large-scale promotion and application.
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Figure CN119242530B_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to the technical field of microorganisms, and in particular to a microbial compound bacterial agent and a preparation method and application thereof. Background Art
[0002] tomato( Solanum lycopersicum L.) was originally a wild berry in South America. It is an annual herbaceous plant of the Solanaceae family and the genus Solanum. It is rich in nutrients such as carotene, vitamin C and B vitamins. It can be used as both a vegetable and a fruit. my country began to grow tomatoes in the 1950s. Tomatoes are cultivated in both the north and south of China and are one of the most commonly cultivated fruits and vegetables in the world.
[0003] Tomatoes are thermophilic crops with wide adaptability to soils, but they require a large amount of fertilizers; they have poor waterlogging resistance and require good drainage conditions. At present, tomato production is mainly based on facility cultivation, but years of facility cultivation have led to serious obstacles to continuous cropping of tomatoes, with long-term continuous cropping and large amounts of pesticides and fertilizers being used. The applied fertilizers will cause an excess of nutrients in the soil and then be converted into insoluble salt substances in the soil, eventually leading to serious soil salinization, which restricts the development of agricultural production.
[0004] In recent years, the concept of green agriculture has been increasingly accepted by the public. Since some microorganisms have relatively single functions, with the continuous deepening of the development of functional microorganisms, researchers have found that the control effect of a combination of multiple microbial agents is higher than that of a single microbial agent, and the effect is more stable. If multiple microbial agents are developed for composite use, combined with the advantages of multiple functional microorganisms, it may be possible to synergistically improve the soil environment and promote plant growth, which is also a green way to solve soil problems. Summary of the invention
[0005] The invention provides a microbial compound agent which can be used to alleviate the problem of continuous cropping obstacles in tomato planting.
[0006] The technical solution of the present invention is as follows:
[0007] A microbial compound agent, comprising Bacillus spores from Saroma Lake ( Sporosarcina saromensis ) ZIP-102 and Serratia marcescens ( Serratia marcescens )GRSB-147; of which:
[0008] The Saroma Lake Sporosarcina ZIP-102 was deposited on October 9, 2024 at the General Microbiological Center of China Microbiological Culture Collection Administration, address: No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, with a deposit number of CGMCC No. 32154;
[0009] The deposit number of the Serratia marcescens GRSB-147 is CGMCC No.26994.
[0010] Preferably, the bacterial concentration of the microbial composite agent is (0.2-0.4)×10 9 CFU / mL.
[0011] The preparation method of the microbial composite bacterial agent is to mix the fermentation liquid of ZIP-102 of Zaroma sporosarcina with the fermentation liquid of Serratia marcescens GRSB-147 to obtain the compound microbial agent.
[0012] Preferably, the volume ratio between the fermentation broth of Bacillus sporesarcinia ZIP-102 and the fermentation broth of Serratia marcescens GRSB-147 in the microbial composite bacterial agent is 1:(0.5-2).
[0013] Preferably, the preparation method of the fermentation broth of Bacillus sporesarcinia ZIP-102 is:
[0014] (1) activating and culturing the Saroma lacustrine sporosarcina ZIP-102 on a solid culture medium to obtain an activated strain;
[0015] (2) inoculating the activated strain obtained in step (1) into a seed liquid culture medium for culturing to obtain a seed liquid;
[0016] (3) The seed liquid obtained in step (2) is inoculated into a fermentation liquid culture medium and cultured to obtain a fermentation liquid.
[0017] Further preferably, the solid culture medium in step (1) is NA solid culture medium, and the activation culture conditions are activation culture at 37±2°C for 24-30h.
[0018] Further preferably, the seed liquid culture medium in step (2) is NA liquid culture medium, and the culture conditions are 37±2°C and 180~200rpm for 18~25h.
[0019] Further preferably, the fermentation liquid medium in step (3) is NA liquid medium, the seed liquid is inoculated at an inoculation rate of 5-10% of the volume percentage of the fermentation liquid medium, and the culture conditions are 37±2°C and 180-200rpm for 24-30h.
[0020] Preferably, the preparation method of the Serratia marcescens GRSB-147 fermentation broth is:
[0021] (1) activating and culturing Serratia marcescens GRSB-147 on a solid culture medium to obtain an activated strain;
[0022] (2) inoculating the activated strain obtained in step (1) into a seed liquid culture medium for culturing to obtain a seed liquid;
[0023] (3) The seed liquid obtained in step (2) is inoculated into a fermentation liquid culture medium and cultured to obtain a fermentation liquid.
[0024] Further preferably, the solid culture medium in step (1) is NA solid culture medium, and the activation culture conditions are activation culture at 37±2°C for 24-30h.
[0025] Further preferably, the seed liquid culture medium in step (2) is NA liquid culture medium, and the culture conditions are 37±2°C and 180~200rpm for 18~25h.
[0026] Further preferably, the fermentation liquid medium in step (3) is NA liquid medium, the seed liquid is inoculated at an inoculation rate of 5-10% of the volume percentage of the fermentation liquid medium, and the culture conditions are 37±2°C and 180-200rpm for 24-30h.
[0027] The application of the microbial composite agent in tomato planting.
[0028] Preferably, the application method is to apply the microbial compound agent to the soil at an application rate of (0.3-0.4)×10 6 CFU / g soil.
[0029] Beneficial effects of the present invention:
[0030] The microbial composite agent of the present invention can effectively improve the soil environment when applied during the tomato planting period. Compared with a single agent, the application of the composite agent of the present invention can increase the tomato plant height by about 4.25%, the tomato stem diameter by about 8.67%, and the tomato fresh weight by about 5.68%, thereby promoting tomato growth and reducing the use of chemical fertilizers. It has a development prospect of large-scale promotion and application. BRIEF DESCRIPTION OF THE DRAWINGS
[0031] Figure 1 This is a picture of strain ZIP-102 observed under a microscope after Gram staining;
[0032] Figure 2 This is a picture of the siderophore production effect of strain ZIP-102;
[0033] Figure 3 This is a picture of the nitrogen fixation effect of strain ZIP-102;
[0034] Figure 4 This is a picture of the organophosphate decomposition effect of strain GRSB-147;
[0035] Figure 5 This is a picture of the inorganic phosphorus decomposition effect of strain GRSB-147;
[0036] Figure 6 This is a picture of the protease production effect of strain ZIP-102;
[0037] Figure 7 This is a picture of the amylase production effect of strain ZIP-102;
[0038] Figure 8 This is a picture of the lipase production effect of strain ZIP-102;
[0039] Fig. 9 This is a picture of the cellulase production effect of strain ZIP-102. DETAILED DESCRIPTION
[0040] The following is explained in conjunction with specific embodiments:
[0041] Source of experimental materials:
[0042] Serratia marcescens ( Serratia marcescens )GRSB-147: The deposit number is: CGMCC No.26994.
[0043] Example 1: Isolation, screening and identification of Bacillus sporosarcina ZIP-102
[0044] A bacterial strain was isolated and screened from the tomato rhizosphere soil in Qingzhou, Shandong, China, and we named it "ZIP-102".
[0045] The above-screened strain ZIP-102 was observed under a microscope after Gram staining. Figure 1 As shown, the bacteria are short rod-shaped; Gram stain is purple, and they are Gram-positive bacteria.
[0046] The 16S rDNA sequence of the strain ZIP-102 obtained by the above screening was sequenced, and the sequencing result is shown in SEQ ID NO.1; the obtained 16S rDNA sequence was subjected to BLAST analysis with the existing sequences in the NCBI database, and strains with similar homology were selected, and the phylogenetic tree was constructed using MEGA X software, and the construction method was Neighbor-joining. The results showed that the strain ZIP-102 obtained by the above screening had a similar structure to the sporosarcina sarcoides ( Sporosarcina saromensis ) is 82% similar to the strain, and the evolutionary distance is relatively close. Combined with the physiological and biochemical characteristics of the strain, it was identified as Spore Sarcinia sarcoides ( Sporosarcina saromensis ).
[0047] Sporosarcina sarcoides Sporosarcina saromensis) ZIP-102, deposited on October 9, 2024 in the General Microbiology Center of China Microorganism Culture Collection Administration, address: No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, with the deposit number CGMCC No.32154.
[0048] Example 2: Evaluation of the growth-promoting function of Bacillus sporosarcina ZIP-102 and Serratia marcescens GRSB-147
[0049] 1. Bacteria activation:
[0050] Sporosarcina ZIP-102 and Serratia marcescens GRSB-147 were streaked onto NA solid culture medium, respectively, and cultured at 37°C for 24 hours to grow single colonies, i.e., activated strains.
[0051] Among them, NA solid culture medium includes the following components: peptone 10.0g / L, beef extract 3.0g / L, sodium chloride 5.0g / L, agar 15.0g / L, pH 7.0~7.5.
[0052] 2. Evaluation of growth-promoting function:
[0053] ①Ability to produce iron carriers:
[0054] Use a sterilized toothpick to pick up the activated bacteria and inoculate them in the CAS detection medium, and culture them at 28°C for 3 days. After the culture is completed, use a vernier caliper to measure the diameter of the transparent circle (D) and the colony diameter (d) in the CAS detection medium.
[0055] The presence of chlorazuron and hexadecyltrimethylammonium bromide in CAS medium will react with Fe 3+ The iron carrier produced by the bacteria combines with the chelate and takes away the Fe in the chelate. 3+ , which causes the color of the culture medium around the bacteria to change. Therefore, the iron carrier production ability of the tested bacteria can be evaluated based on the D / d value.
[0056] Among them, the components of CAS detection culture medium are as follows: chrome azuro blue S 60.5 mg / L, hexadecyltrimethylammonium bromide 72.9 mg / L, ferric chloride hexahydrate 2.645 mg / L, sodium dihydrogen phosphate dihydrate 295.25 mg / L, disodium hydrogen phosphate dodecahydrate 1213.5 mg / L, ammonium chloride 62.5 mg / L, potassium dihydrogen phosphate 37.5 mg / L, sodium chloride 62.5 mg / L, agar 9000 mg / L, pH 6.7~6.9; high pressure sterilization at 116°C for 30 min.
[0057] Photos after the cultivation of Lake Saroma Sporosarcina ZIP-102 Figure 2 As shown. Figure 2It can be seen that Bacillus sporosarcina ZIP-102 has the ability to produce siderophore. The specific siderophore production function evaluation results are shown in Table 1.
[0058] ②Nitrogen fixation ability:
[0059] Use a sterile inoculation loop to pick up the activated bacteria and inoculate them into the Axubei nitrogen-free solid culture medium, and culture them at 28°C for 7 days. After the culture is completed, observe whether there are colonies in the culture medium. If there are colonies, it means that the strain has the ability to fix nitrogen.
[0060] Among them, the components of Axubei nitrogen-free solid culture medium are as follows: 10g glucose, 0.2g dipotassium hydrogen phosphate, 0.2g magnesium sulfate, 0.2g sodium chloride, 0.2g calcium sulfate, 5g calcium carbonate, 1000mL distilled water, 20g agar, pH 7.0~7.2, high pressure sterilization at 116℃ for 30min.
[0061] Photos after the cultivation of Lake Saroma Sporosarcina ZIP-102 Figure 3 As shown. Figure 3 It can be seen that colonies were formed in the nitrogen-free solid culture medium of Asubei, which indicates that the Saroma Lake Sporosarcina ZIP-102 has the ability to fix nitrogen.
[0062] ③ Phosphorus solubilization capacity
[0063] Use a sterilized toothpick to pick up the activated bacteria and inoculate them into organic phosphorus culture medium and inorganic phosphorus culture medium respectively, and culture them at 28℃ for 7 days. Observe the size of the degradation circle, and use a vernier caliper to measure the diameter of the transparent circle (D) and the colony diameter (d). The phosphate solubilization ability of the strain is evaluated according to the phosphate solubilization coefficient (i.e., D / d value).
[0064] The photo of Serratia marcescens GRSB-147 after cultivation in organophosphate medium Figure 4 The photos after the cultivation in inorganic phosphorus medium are shown in Figure 5 As shown. Figures 4 and 5 It can be seen that the strain has degradation zones in both organic phosphorus medium and inorganic phosphorus medium, which indicates that Serratia marcescens GRSB-147 has the ability to degrade both organic and inorganic phosphorus.
[0065] Among them, the components of the organophosphorus culture medium are as follows: glucose 10.0g, ammonium sulfate 0.5g, sodium chloride 0.3g, magnesium sulfate 0.3g, manganese sulfate 0.03g, sulfuric acid 0.3g, ferrous sulfate 0.03g, calcium phosphate 5.0g, lecithin 0.2g, agar 15.0g, water 1000mL, pH 7.0~7.5, high pressure sterilization at 116℃ for 30min.
[0066] Among them, the components of the inorganic phosphorus culture medium are as follows: glucose 10.0g, ammonium sulfate 0.5g, sodium chloride 0.3g, magnesium sulfate 0.3g, manganese sulfate 0.03g, potassium sulfate 0.3g, ferrous sulfate 0.03g, calcium phosphate 5.0g, agar 15.0g, water 1000mL, pH 7.0~7.5, high pressure sterilization at 116℃ for 30min.
[0067] Table 1. Evaluation results of growth-promoting function of ZIP-102 and GRSB-147
[0068]
[0069] Note: “*” represents the strain has the growth-promoting function, “ / ” represents the strain does not have the growth-promoting function.
[0070] As shown in Table 1, Bacillus sporosarcina ZIP-102 has the ability to produce siderophores and fix nitrogen; and Serratia marcescens GRSB-147 has the ability to degrade organic and inorganic phosphorus.
[0071] Example 3: Evaluation of enzyme production ability of Bacillus sporesarcinus ZIP-102
[0072] 1. Bacteria activation:
[0073] Streak the Saroma Lake Sporosarcina ZIP-102 on NA solid culture medium and culture at 37°C for 24 hours to grow a single colony, i.e., the activated strain.
[0074] 2. Evaluation of the protease production ability of bacterial strains:
[0075] Use a sterilized toothpick to pick up the activated bacteria and inoculate them in a skimmed milk powder culture medium, and culture them at 28°C for 5 days. After the culture is completed, observe the color change of the culture medium around the bacteria, and use a vernier caliper to measure the diameter of the transparent circle in the culture medium (D) and the colony diameter (d), and calculate the D / d value.
[0076] The proteins present in the skimmed milk powder culture medium can be broken down into small molecular amino acids and peptides by the proteases produced by the bacteria, causing the color of the culture medium surrounding the bacteria to change from yellow-white to transparent. Therefore, the protease production ability of the tested bacteria can be evaluated by observing the color changes of the culture medium surrounding the bacteria and the D / d value.
[0077] The components of the skimmed milk powder culture medium are as follows: 0.5 g beef extract, 1 g peptone, 0.5 g NaCl, 3 g skimmed milk powder, 2 g agar, 100 mL distilled water, pH 7.0-7.2.
[0078] Photos after the training Figure 6 shown by Figure 6It can be seen that the color of the culture medium around the bacteria changed from yellow-white to transparent, which indicates that Sporosarcina ZIP-102 has the ability to produce protease.
[0079] 3. Evaluation of the ability of bacterial strains to produce amylase:
[0080] Use a sterilized toothpick to pick up the activated bacteria and inoculate them in starch culture medium, and culture them at 28°C for 5 days. After the culture, add Lugol's iodine solution, observe the color change of the culture medium around the bacteria, and use a vernier caliper to measure the diameter of the transparent circle in the culture medium (D) and the colony diameter (d), and calculate the D / d value.
[0081] The starch present in the starch culture medium can be decomposed into small molecule sugars such as maltose and glucose by the amylase produced by the bacteria, and the starch turns blue when it meets Lugol's iodine solution. When the starch around the bacteria is enzymatically hydrolyzed, the color of the culture medium around the bacteria will change from blue-purple to transparent. Therefore, the amylase production ability of the tested bacteria can be evaluated by observing the color changes of the culture medium around the bacteria and the D / d value.
[0082] The components of the starch culture medium are as follows: 2 g soluble starch, 10 g peptone, 5 g beef extract, 5 g sodium chloride, 20 g agar, pH 7.2, dilute to 1 L with distilled water; sterilize at 121°C for 30 min.
[0083] Among them, the components of Lugol's iodine solution are as follows: 1g iodine tablets, 2g potassium iodide, and 300mL distilled water.
[0084] Photos after the training Figure 7 shown by Figure 7 It can be seen that the color of the culture medium around the bacteria changed from blue-purple to transparent, which indicates that Sporosarcina ZIP-102 has the ability to produce amylase.
[0085] 4. Evaluation of Lipase Production Capacity of Bacteria:
[0086] Use a sterilized toothpick to pick up the activated bacteria and inoculate them in the fat culture medium, and culture them at 28°C for 5 days. After the culture, observe the color change of the culture medium around the bacteria, and use a vernier caliper to measure the diameter of the transparent circle in the culture medium (D) and the diameter of the colony (d), and calculate the D / d value.
[0087] Tributyrin present in the fat culture medium can be decomposed into fatty acids and glycerol by the lipase produced by the bacteria, causing the color of the culture medium surrounding the bacteria to change from white to transparent. Therefore, the lipase-producing ability of the tested bacteria can be evaluated by observing the color change of the culture medium surrounding the bacteria and the D / d value.
[0088] The components of the fat culture medium are as follows: peptone 10 g, yeast powder 5 g, NaCl 10 g, tributyrin 2 mL, agar 20 g, distilled water 1 L, pH 7.5.
[0089] Photos after the training Figure 8 shown by Figure 8 It can be seen that the color of the culture medium around the bacteria changed from white to transparent, which indicates that Sporosarcina ZIP-102 has the ability to produce lipase.
[0090] 5. Evaluation of the ability of bacterial strains to produce cellulase:
[0091] Use a sterilized toothpick to pick up the activated bacteria and inoculate them in the cellulose culture medium, and culture them at 28°C for 5 days. After the culture is completed, observe the color change of the culture medium around the bacteria, and use a vernier caliper to measure the diameter of the transparent circle in the culture medium (D) and the colony diameter (d), and calculate the D / d value.
[0092] The sodium carboxymethyl cellulose present in the cellulose culture medium changes from colorless to red when stained with Congo red dye, and the sodium carboxymethyl cellulose can be decomposed by the cellulase produced by the bacteria, causing the color of the culture medium around the bacteria to change from red to transparent. Therefore, the cellulase production ability of the tested bacteria can be evaluated by observing the color change of the culture medium around the bacteria and the D / d value.
[0093] The components of the cellulose culture medium are as follows: 10 g of peptone, 5 g of yeast powder, 10 g of sodium chloride, 10 g of sodium carboxymethyl cellulose, 20 g of agar, and 1 L of distilled water.
[0094] Photos after the training Fig. 9 shown by Fig. 9 It can be seen that the color of the culture medium around the bacteria changed from red to transparent, which indicates that Sporosarcina ZIP-102 has the ability to produce cellulase.
[0095] The measurement results of D / d values in the above experiments are shown in Table 2 below:
[0096] Table 2. Evaluation results of enzyme production capacity of ZIP-102
[0097]
[0098] As shown in Table 2, Bacillus sporosarcina ZIP-102 has good ability to produce protease, amylase, lipase and cellulase.
[0099] Example 4: Preparation of microbial compound agent
[0100] 1. Preparation of fermentation broth of Bacillus sporesarcinia ZIP-102 from Saroma Lake:
[0101] (1) Activate and culture the ZIP-102 of Bacillus spore-forming bacteria on NA solid medium at 37°C for 24 hours to obtain an activated strain;
[0102] (2) inoculating the activated strain obtained in step (1) into NA liquid culture medium, and culturing at 37° C. and 180 rpm for 18 h to obtain seed solution;
[0103] (3) The seed solution prepared in step (2) was inoculated into a new NA liquid culture medium at a volume percentage of 5%, and cultured at 37°C and 180 rpm for 24 h to obtain a fermentation broth with a bacterial cell concentration of 0.4×10 9 CFU / mL.
[0104] 2. Preparation of Serratia marcescens GRSB-147 fermentation broth:
[0105] (1) Activating Serratia marcescens GRSB-147 on NA solid medium at 37°C for 24 hours to obtain an activated strain;
[0106] (2) inoculating the activated strain obtained in step (1) into NA liquid culture medium, and culturing at 37° C. and 180 rpm for 18 h to obtain seed solution;
[0107] (3) The seed solution prepared in step (2) was inoculated into a new NA liquid culture medium at a volume percentage of 5%, and cultured at 37°C and 180 rpm for 24 h to obtain a fermentation broth with a bacterial cell concentration of 0.3×10 9 CFU / mL.
[0108] 3. Preparation of microbial compound agents:
[0109] 1:1 compound bacterial agent: The fermentation liquid of the above-mentioned ZIP-102 sporosarcina and the fermentation liquid of the above-mentioned Serratia marcescens GRSB-147 were compounded in a volume ratio of 1:1, and the bacterial concentration was 0.35×10 9 CFU / mL.
[0110] 1:2 compound bacterial agent: The fermentation liquid of the above-mentioned ZIP-102 sporogenous Sarcina sarcoides and the fermentation liquid of the above-mentioned Serratia marcescens GRSB-147 were compounded in a volume ratio of 1:2, and the bacterial concentration was 0.33×10 9 CFU / mL.
[0111] 2:1 compound bacterial agent: The fermentation liquid of the above-mentioned ZIP-102 sporosarcina and the fermentation liquid of the above-mentioned Serratia marcescens GRSB-147 were compounded in a volume ratio of 2:1, and the bacterial concentration was 0.37×10 9 CFU / mL.
[0112] Example 5: Microbial compound agent promotes the growth of tomato plants
[0113] This experiment was conducted in the laboratory. First, 60 pots of healthy, pest-free, and three-true-leaf tomato seedlings of the Provence variety were selected and planted in pots. The mass ratio of soil (arable soil) to vermiculite (to increase the looseness and air permeability of the soil) in the pots was 1:1, and the total weight was 1.5±0.1kg.
[0114] The ZIP-102 group, GRSB-147 group, 1:1 compound bacterial agent group, 1:2 compound bacterial agent group, 2:1 compound bacterial agent group and control group were set up respectively, with 30 pots in each group; among them, the ZIP-102 group was treated with the fermentation liquid of ZIP-102 prepared in Example 4, and the application amount was 0.35×10 6 CFU / g; GRSB-124 group was treated with the fermentation broth of Serratia marcescens GRSB-147 prepared in Example 4, with an application amount of 0.35×10 6 CFU / g; the 1:1 compound bacterial agent group, the 1:2 compound bacterial agent group, and the 2:1 compound bacterial agent group were respectively applied with the compound bacterial agent prepared in Example 4, and the application amount was 0.35×10 6 CFU / g; the control group was treated with the same volume of sterile PBS buffer as the 1:1 compound bacterial agent group. The above 6 groups of tomato seedlings were cultured in the same environment, and the experimental period was 25 days.
[0115] After the experiment, the growth data of the above six groups of tomato plants, such as plant height, stem diameter, dry weight, fresh weight and leaf chlorophyll content, were measured. The method for measuring leaf chlorophyll was as follows: 1g of fresh leaves were placed in liquid nitrogen for low temperature treatment for 20s, then ground and dissolved in a mortar with 20mL of anhydrous ethanol, and the absorbance was measured at 645nm and 663nm using an ultraviolet spectrophotometer. The chlorophyll content (C T ), the calculation formula is:
[0116] C a (mg / L)=12.7OD 663 -2.69OD 645 ;
[0117] C b (mg / L)=22.9OD 645 -4.68OD 663 ;
[0118] C T (mg / L)=C a +C b .
[0119] The measurement results are shown in Table 3 below:
[0120] Table 3. Growth-promoting effect of microbial compound agents on tomato plants
[0121]
[0122] As shown in Table 3, compared with the control group, the addition of bacterial agents can promote the growth of tomato plants to varying degrees. Specifically, compared with the control group, the application of 1:1 compound bacterial agents increased the plant height by about 23.09%, the stem thickness by about 17.73%, the dry weight by about 53.12%, the fresh weight by about 48.77%, and the chlorophyll by about 0.87%. Compared with a single bacterial agent, the application of 1:1 compound bacterial agents can significantly increase the plant height, stem thickness, and fresh weight of tomato plants. Specifically, compared with ZIP-102, the application of 1:1 compound bacterial agents increased the plant height by about 4.25%, the stem thickness by about 8.67%, and the fresh weight by about 5.68%.
Claims
1. A microbial compound agent, characterized in that: Including Bacillus sporosarcina ( Sporosarcina saromensis ) ZIP-102 and Serratia marcescens ( Serratia marcescens )GRSB-147; of which: The Saroma Lake Sporosarcina ZIP-102 was deposited on October 9, 2024 at the General Microbiological Center of China National Microbiological Culture Collection Administration, address: No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, with a deposit number of CGMCC No. 32154; The deposit number of the Serratia marcescens GRSB-147 is CGMCC No.26994.
2. The microbial composite agent according to claim 1, characterized in that: The bacterial concentration of the microbial composite agent is (0.2~0.4)×10 9 CFU / mL.
3. The method for preparing the microbial composite agent according to claim 1, characterized in that: The fermentation liquid of the Saroma lacustrine sporosarcina ZIP-102 is mixed with the fermentation liquid of the Serratia marcescens GRSB-147.
4. The preparation method according to claim 3, characterized in that: The volume ratio between the fermentation broth of the Saroma lacustrine sporosarcina ZIP-102 and the fermentation broth of the Serratia marcescens GRSB-147 is 1:(0.5-2).
5. The preparation method according to claim 3, characterized in that: The preparation method of the fermentation liquid of the Zoroma Lake Sporosarcina ZIP-102 is as follows: (1) activating and culturing the Saroma lacustrine sporosarcina ZIP-102 on a solid culture medium to obtain an activated strain; (2) inoculating the activated strain obtained in step (1) into a seed liquid culture medium for culturing to obtain a seed liquid; (3) The seed liquid obtained in step (2) is inoculated into a fermentation liquid culture medium for culturing to obtain a fermentation liquid.
6. The preparation method according to claim 3, characterized in that: The preparation method of the fermentation broth of Serratia marcescens GRSB-147 is as follows: (1) activating and culturing Serratia marcescens GRSB-147 on a solid culture medium to obtain an activated strain; (2) inoculating the activated strain obtained in step (1) into a seed liquid culture medium for culturing to obtain a seed liquid; (3) The seed liquid obtained in step (2) is inoculated into a fermentation liquid culture medium and cultured to obtain a fermentation liquid.
7. The preparation method according to any one of claims 5 to 6, characterized in that The solid culture medium is a NA solid culture medium; the seed liquid culture medium or the fermentation liquid culture medium is a NA liquid culture medium.
8. The use of the microbial compound agent according to claim 1, characterized in that: Used in tomato cultivation.
9. The use according to claim 8, characterized in that The application method is to apply the microbial compound agent to the soil at an application rate of (0.3-0.4)×10 6 CFU / g soil.