CN107058151B - Microbial bacterium with fermentation water retention function and application thereof - Google Patents
Microbial bacterium with fermentation water retention function and application thereof Download PDFInfo
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Abstract
The invention discloses a microbial strain with fermentation water retention function and application thereof, wherein a bacterial strain MTZ102 is a bacillus amyloliquefaciens with the functions of producing fatty acid, organic acid, capsular polysaccharide and the like through fermentation, and can produce secondary metabolites under different fermentation conditions. The strain MTZ102 is derived from drought-barren corn rhizosphere soil, can obviously promote the tolerance of the crops such as the crowndaisy chrysanthemum, the corn and the like to the low water content of the soil, and provides an excellent liquid bacterial fertilizer for the growth of the crops such as the crowndaisy chrysanthemum, the corn and the like in the relatively drought soil.
Description
Technical Field
The invention relates to the field of microbial organic fertilizers of microorganisms, and particularly relates to a microorganism with a fermentation water retention function and application thereof.
Background
Drought is one of the most serious disasters in the world, and the occurrence frequency, duration, influence range, caused loss and the like of the drought are at the top of various natural disasters and far exceed other meteorological disasters. In 2014, crops in the whole country have 3.4 million mu of drought area, 1.8 million mu of disaster area, 8516 million mu of disaster area and 2227 million mu of absolute area, and grain loss 2006 million tons, economic crop loss 276 million yuan and direct economic total loss 910 million yuan are caused by drought.
For the last 40 years. With the increase of the content of greenhouse gases in the atmosphere, the climate in China generally has a tendency of warming and drying, and the tendency causes great adverse effects on agriculture in China. Research shows that the method is not changed in the existing planting system, planting variety and production level. 2030 and 2050, the food production potential of China can be reduced by about 10% due to the extreme climatic events caused by climate change. Wherein 3 large crops of wheat, rice and corn are mainly yield-reduced, have certain influence on quality, and will seriously influence the grain safety of China. Therefore, the method reduces the influence and the response measures of drought on agricultural production in China, is extremely important for developing sustainable agriculture, and has practical and profound significance. Therefore, the water and soil loss resistance, the drought resistance and the water conservation become the problems that whether agriculture in China can be continuously developed in the future or not needs to be urgently solved.
The water retention agent is called a Super Absorbent Polymer (SAP), which is a compound capable of absorbing water several times its original size and weight. The hydrogel formed by the water-retaining agent after absorbing water can slowly release water for crops to utilize, and simultaneously can enhance the water-retaining property of soil, improve the soil structure, influence the physical and chemical properties of the soil, the growth and development of crop roots and the like, further influence the migration rule of water and solute in the soil and the physiological function of the crops, and improve the utilization rate of the fertilizer. At present, water-retaining agents are used in a plurality of fields such as industry, agriculture, building, gardening, sanitation and the like in nearly 30 countries, are well developed in agriculture, and have wide application prospects. However, environmental problems have begun to occur from the use of the first generation water retention agents. The first generation water retention agent uses acrylonitrile monomer as raw material, and the residual raw material can generate toxicity, thus affecting the environmental safety. The second generation starch-polyacrylic acid graft copolymer developed in 1975 by the japan ocean corporation, although having low cost and toxicity, has low thermal stability and poor salt tolerance, and has a reduced water absorption property in high-salt soil and causes severe soil hardening. The novel water-retaining agent developed in the middle of the 80 s is improved in salt resistance, heat stability, water retention and the like, and the problem of soil hardening is effectively improved. However, the environmental problems possibly caused by the blind popularization and use of the water-retaining agent in the application process make the evaluation of the agricultural safety inevitable. Although the water retention agents developed and used at present have no obvious influence on the physicochemical properties of the soil, with the progress of modern technology, the evaluation of the basic physicochemical properties (including water retention performance) of the soil is far from enough. In particular, soil microorganisms are an important component of the soil ecosystem and are closely related to soil material circulation and energy flow. The study of the ecological impact of water-retaining agents on soil microorganisms is still blank (lish et al, 2012). And metabolic products such as fatty acid, organic acid, capsular polysaccharide and the like generated by fermentation of beneficial microorganisms can form a protective film in capillary gaps in soil, so that evaporation of water in the gaps of the soil can be retarded, and the purposes of water retention and drought resistance of the soil are achieved.
The bacillus amyloliquefaciens is a bacterium with high affinity with bacillus subtilis, and can generate a series of metabolites in the growth process of the bacillus amyloliquefaciens, so that the bacillus amyloliquefaciens can widely inhibit the activity of fungi and bacteria, and can also generate fatty acid, organic acid, capsular polysaccharide and the like to form a biofilm substance to be suspended on the surface of water. Therefore, in recent years, a large number of researchers have conducted extensive research on the novel discovery and application of Bacillus amyloliquefaciens, gene function research, and optimization of fermentation conditions. The bacillus amyloliquefaciens can generate a plurality of antibacterial substances, has wide bactericidal spectrum and has prevention and treatment effects on pathogenic bacteria, so the bacillus amyloliquefaciens has very obvious effect on preventing and treating diseases of crops, vegetables and melons and fruits, and simultaneously provides an effective way for reducing chemical pesticides and ensuring food safety. More importantly, the research finds that the bacillus amyloliquefaciens can generate a high-molecular biological ester film when fermenting certain specific organic matters, and the high-molecular biological ester film is suspended on the surface of water, so that the evaporation of water molecules can be reduced, and the purposes of water retention and drought resistance are achieved.
Disclosure of Invention
Aiming at the problems, the invention aims to provide a microbial strain with the functions of fermentation and water retention, which has the advantages of high growth speed, strong environmental adaptability and strong stress tolerance, and the microbial strain can remarkably improve the yield and the quality of vegetables and crops by suspending a macromolecular biological ester film generated by fermenting organic matters through a strain on the surface of water molecules in capillary pores of soil, reducing the water evaporation of the soil and improving the water content of the soil.
In order to realize the technical purpose, the scheme of the invention is as follows: a microorganism strain with fermentation and water retention functions comprises a strain MTZ102 which is Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) with the collection number of GDMCC 60099 and the collection date of 2016, 10 and 31 days;
the morphology and physiological and biochemical characteristics of the strain MTZ102 are as follows:
the bacterial strain MTZ102 is gram-positive bacteria, and the cells are rod-shaped and can produce spores;
the growth speed on an LB agar plate is high, the colony is circular, the middle of the colony is small and convex (growing for 36 hours at 37 ℃), the colony is milky and opaque, and the diameter of the colony is 2-8 mm; the cell size is 2.7-4.1x1.5-1.7 μm, and the cell is rod-shaped and has terminal flagellum; the optimal growth temperature is 27-40 ℃, the pH growth range is 4-10, and the optimal growth pH value is 6-8;
facultative anaerobes capable of growing on media with D-galactose, L-arabitol, sucrose, dextrin, sorbose, D-fructose, D-glucose, cellobiose, turanose, trehalose, mannitol, D-arabitol, sorbitol, hexitol, melezitose, galactitol, sorbitol, L-fructose, sodium gluconate, sodium lactate, sodium succinate, malic acid, glycerol, soybean oil, sodium D-glucuronate, succimide, N-acetylglucosamine, L-glutamic acid, L-alanine, L-glutamine, L-histidine, L-cystine, aspartic acid as the sole carbon source;
the growth range of the pH is wide, and the pH can be resistant to 4-10; the NaCl concentration is lower than 2.0 percent, and the growth can be realized; can tolerate streptomycin, ampicillin and tetracycline of 300 mug/mL and gentamicin and erythromycin of 100 mug/mL. The growth of the streptomycin, the chloramphenicol and the neomycin can be inhibited under the concentration condition of more than 50 mug/mL; produce siderophore, the indole reaction is positive, and can secrete auxin IAA.
Microbial organic fertilizer using bacterial strain MTZ102, wherein the content of bacterial strain MTZ102 is 108The mass ratio of total nitrogen to total nitrogen is 4-5%, the mass ratio of total nitrogen to total nitrogen is more than 90% of organic nitrogen, the mass ratio of total nitrogen, phosphorus and potassium nutrients is 6-10%, and the mass ratio of organic matter content is not less than 45%.
A method for producing microbial organic fertilizer of a strain MTZ102,
the first step, inoculating the strain MTZ102 into a culture solution of modified MRS type for fermentation production, wherein the conditions of the fermentation production are as follows: the culture temperature is 28-35 ℃, the stirring speed is 180-300 rpm, spores are formed in the middle and later stages of fermentation, and the bacteria or spore content in the fermentation liquor is more than or equal to 1 multiplied by 109cfu·ml-1；
Secondly, performing solid compost fermentation on the bacterial strain MTZ102 and the decomposed agricultural organic waste mixture, adding 200L of zymocyte liquid into each ton of solid organic materials, turning the mixture for 1 time every 7 to 12 days in the solid fermentation process, stopping the solid fermentation after the solid fermentation temperature is not more than 80 ℃, and enabling the content of zymocyte to reach 1 multiplied by 10 after the solid fermentation is generally finished for 25 to 40 days8cfu·g-1Thus, a microbial organic fertilizer containing a mixture of the fermentation tubes MTZ102 and agricultural organic waste is obtained.
Preferably, after the microbial organic fertilizer in the second step is decomposed, the mass ratio content of organic matters is more than or equal to 35%, the mass ratio content of organic nitrogen is 1.2-2%, and the mass ratio of water content is more than 25%.
Preferably, the ratio of the culture solution in the first step is as follows: 0.15-0.25 g of yeast powder, 6-14 g of molasses and KH2PO40.01～0.02g，K2HPO40.01-0.02 g, 3-9 g of soybean meal, 0.5-1.5 g of wheat bran, and adding water to 1L, wherein the pH value is 6-8; after the preparation of the culture solution, the culture solution is sterilized at 121 ℃ for 20 min.
Preferably, in the second step, the total nitrogen content of the microbial organic fertilizer is 4-5% by mass, more than 90% by mass of the total nitrogen is organic nitrogen, the total nitrogen, phosphorus and potassium nutrient content is 6-10% by mass, and the organic matter content is not less than 45% by mass.
Preferably, the agricultural organic waste in the second step is one or a combination of straws, bran coats, bean pulp and wheat bran.
The application of the microbial organic fertilizer of the strain MTZ102 in the cultivation of the crowndaisy chrysanthemum and the corn.
The invention has the beneficial effects that 1, the bacterial strain MTZ102 is a bacillus amyloliquefaciens with the functions of producing fatty acid, organic acid, capsular polysaccharide and the like through fermentation, the secondary metabolites can be produced under different fermentation conditions, and compared with the artificially synthesized water-retaining agent, the secondary metabolites produced through microbial fermentation can be biodegraded, are environment-friendly and safe, and cannot cause harm and secondary pollution to the soil environment.
2. The strain MTZ102 is derived from drought-barren corn rhizosphere soil, can obviously promote the tolerance of the crops such as the crowndaisy chrysanthemum, the corn and the like to the low water content of the soil, and provides an excellent liquid bacterial fertilizer for the growth of the crops such as the crowndaisy chrysanthemum, the corn and the like in the relatively drought soil.
3. The microbial organic fertilizer prepared by the strain MTZ102 contains an organic micromolecular film which retards water volatilization, keeps the water content of soil, can obviously improve the absorption of crops such as crowndaisy chrysanthemum, corn and the like on mineral substances and organic nutrients, ensures the chlorophyll content, the Vc content of leaves and the yield of the leaf tissues of the crops, and has good effects of increasing the quality and the yield.
Detailed Description
The present invention will be described in further detail with reference to specific examples.
The microbial strain with the functions of producing fatty acid, organic acid, capsular polysaccharide and the like by fermentation kills non-bacillus by adopting 80 ℃, and is cultured by modified MRS (1L): 10g of peptone, 10g of beef extract, 5g of yeast extract, 2.0g of diammonium hydrogen citrate, 20g of glucose, 2g of dipotassium hydrogen phosphate, 1.63g of ammonium sulfate, 5g of sodium acetate, 0.58g of magnesium sulfate, 0.25g of manganese sulfate, 80mL of Tween, 1g of calcium carbonate, 18g of agar, adding distilled water to 1000mL, and carrying out culture under anaerobic and aerobic conditions, wherein the pH value is 6.4 and 0.2 respectively. 5 bacillus producing capsular polysaccharide are separated and purified from corn plants growing on Guangzhou barren soil. Further screening the capacities of producing fatty acid, organic acid, capsular polysaccharide and the like by fermenting the 5 strains to obtain a strain MTZ102 which has high growth speed and strong capsular polysaccharide producing capacity, can produce capsular polysaccharide under different culture medium formulas and is stable in capsular polysaccharide production; the fermentation liquor of the strain is used as an inoculant, and the growth of crops such as crowndaisy chrysanthemum, corn and the like can be promoted by applying the soil with drought and water shortage.
A microbe with fermenting and water-retaining functions includes bacterial strain MTZ102,
the strain MTZ102 is Bacillus amyloliquefaciens (Bacillus amyloliquefaciens), the preservation number is GDMCC 60099, and the preservation date is 2016, 10 and 31 days; the preservation unit: guangdong province microbial strain preservation center, address: guangzhou city, first furious Zhonglu No. 100 large yard No. 59 building No. 5.
The morphological and physiological and biochemical characteristics of the strain MTZ102 are as follows:
the bacterial strain MTZ102 is gram-positive bacteria, and the cells are rod-shaped and can produce spores;
the growth speed on an LB agar plate is high, the colony is circular, the middle of the colony is small and convex (growing for 36 hours at 37 ℃), the colony is milky and opaque, and the diameter of the colony is 2-8 mm; the cell size is 2.7-4.1x1.5-1.7 μm, and the cell is rod-shaped and has terminal flagellum; the optimal growth temperature is 27-40 ℃, the pH growth range is 4-10, and the optimal growth pH value is 6-8;
facultative anaerobes capable of growing on media with D-galactose, L-arabitol, sucrose, dextrin, sorbose, D-fructose, D-glucose, cellobiose, turanose, trehalose, mannitol, D-arabitol, sorbitol, hexitol, melezitose, galactitol, sorbitol, L-fructose, sodium gluconate, sodium lactate, sodium succinate, malic acid, glycerol, soybean oil, sodium D-glucuronate, succimide, N-acetylglucosamine, L-glutamic acid, L-alanine, L-glutamine, L-histidine, L-cystine, aspartic acid as the sole carbon source;
the growth range of the pH is wide, and the pH can be resistant to 4-10; the NaCl concentration is lower than 2.0 percent, and the growth can be realized; can tolerate streptomycin, ampicillin and tetracycline of 300 mug/mL and gentamicin and erythromycin of 100 mug/mL. The growth of the streptomycin, the chloramphenicol and the neomycin can be inhibited under the concentration condition of more than 50 mug/mL; produce siderophore, the indole reaction is positive, and can secrete auxin IAA.
(II) determination of molecular Classification position of Bacillus amyloliquefaciens MTZ102 of the invention
PCR amplification is carried out by adopting bacterial 16S rDNA gene universal primers 25f and 1492r, forward and backward direction sequence determination is directly carried out on a PCR product, an obtained DNA sequence is input into GenBank for comparison, and the species and the genus position of the bacillus in the taxonomy are preliminarily determined. As a result, it was found that the Bacillus of the present invention belongs to the genus Bacillus (Bacillus sp.) and has 98.3% similarity to the Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) model strain LMG 12234. Physiological and biochemical test characteristics and 16S rRNA gene sequence analysis show that the bacillus amyloliquefaciens MTZ102 and the bacillus amyloliquefaciens model strain LMG12234 have differences. According to the physiological and biochemical characteristics and the 16S rDNA sequence analysis result, the Bacillus amyloliquefaciens disclosed by the invention belongs to Bacillus (Bacillus sp.) and is a new strain of the Bacillus, and has the capability of producing fatty acid, organic acid and capsular polysaccharide through fermentation. The judgment is made in the following documents: borris, R., et al, Relationship of Bacillus amyloliquefaciens clades associated with strains DSM7T and FZB42T a pro posal for Bacillus amyloliquefaciens subsp.amyloliquefaciens subsp.nov.and Bacillus amyloliquefaciens subsp.plantarum subsp.nov.bated on complete genome sequence ligands. int J. Syst ol Microbiol,2011.61(Pt 8) p.1786-801; fan, B., et al, Transcriptomic profiling of Bacillus amyloliquefaciens FZB42 in response to mail root extensions. BMC Microbiol,2012.12: p.116; yuan, J.et al, Plant growth-promoting rhizobacterium strain NJN-6-engineered bio-organic fertilizer depleted Fusarium salt and expressed the growth of banana plants J.agricultural Food Chem. 2013.61(16): p.3774-80.
(III) separation and purification method of the strain of the invention
3.1 enrichment: weighing 30g of corn rhizosphere soil growing in the barren soil into a triangular flask containing 50mL of sterile water, fully oscillating and scattering to prepare suspension, mixing the suspension with the enriched nutrient solution according to the ratio of 1: 2 proportion, placing in a triangular flask, standing in an incubator, culturing at 30 + -2 deg.C for 3 days, carefully taking out supernatant, placing in another sterilized beaker, and heating to 80 deg.C to kill non-spore bacteria.
Taking 500 mu L to 250mL of enriched nutrient solution of the heat treatment bacterial suspension in the first generation culture bottle to carry out second generation culture, wherein the culture method is the same as that of the first generation culture.
3.2, separation and purification: sucking 1ml of second generation thallus culture solution into 9ml sterile water test tube to obtain 10-1Diluting the bacterial liquid by times, and diluting the bacterial suspension with the concentration to 10-2 and 10-3 times by the method until the bacterial suspension is diluted to 10-6And (4) doubling. Respectively suck 10-4、10-5、10-6Three gradient bacterial suspension 100 μ L, sequentially adopting plate coating culture method and pouring anaerobic culture method, culturing at 30 + -2 deg.C, observing different cultureThe method is characterized in that under the growth condition of thalli, colonies with good growth vigor and different characteristics are selected and subjected to repeated subculture by using a plate marking method until the shapes, colors, textures and transparencies of the colonies are consistent. And finally, further observing the form of the strain by simple dyeing and microscopic examination, wherein the uniform length, the uniform width and the uniform dyeing condition are taken as the strain purification standards.
3.3, preservation: and culturing the separated and purified strain on a separation culture medium flat plate and a slant for 2-4 days, and collecting thalli on the flat plate. The fractions were stored in 15% sterile glycerol (-20 ℃) and (-80 ℃).
Wherein the enrichment broth (1L): KH (Perkin Elmer)2PO4 0.5g，K2HPO4 0.6g，MgSO47H2O 0.2.2 g, NaCl 0.2g, yeast extract 0.5g, peptone 5.0g, calcium carbonate 0.5g, add deionized water to 1L), pH is natural, about 7.0. + -. 0.2.
Wherein the isolation medium (1L): 2g of yeast extract, 4g of peptone, 0.5g of calcium carbonate, 17g of agar powder and pH 7.0 +/-0.2.
Secondly, the proportion of the components of the microbial organic fertilizer fermented by the microbial bacteria is as follows:
microbial organic fertilizer using bacterial strain MTZ102, wherein the content of bacterial strain MTZ102 is 108The mass ratio of total nitrogen to total nitrogen is 4-5%, the mass ratio of total nitrogen to total nitrogen is more than 90% of organic nitrogen, the mass ratio of total nitrogen, phosphorus and potassium nutrients is 6-10%, and the mass ratio of organic matter content is not less than 45%.
Thirdly, preparing a microbial organic fertilizer fermented by microbial bacteria:
a method for producing microbial organic fertilizer of a strain MTZ102,
the first step, inoculating the strain MTZ102 into a culture solution of modified MRS type for fermentation production, wherein the conditions of the fermentation production are as follows: the culture temperature is 28-35 ℃, the stirring speed is 180-300 rpm, spores are formed in the middle and later stages of fermentation, and the bacteria or spore content in the fermentation liquor is more than or equal to 1 multiplied by 109cfu·ml-1；
Secondly, performing solid compost fermentation on the bacterial strain MTZ102 and the decomposed agricultural organic waste mixture, and adding 100-2 parts of zymophyte liquid into each ton of solid organic materials00L, turning over the pile for 1 time every 7-12 days in the solid fermentation process, keeping the solid fermentation temperature not more than 80 ℃, and finishing the fermentation after 25-40 days to make the content of zymophyte reach 1 × 108cfu·g-1Thus, a microbial organic fertilizer containing a mixture of the fermentation tubes MTZ102 and agricultural organic waste is obtained.
In order to ensure that the proportion of nitrogen, phosphorus and potassium of the organic fertilizer accords with the growth of plants, in the second step, after the microbial organic fertilizer is decomposed, the mass ratio content of organic matters is more than or equal to 35%, the mass ratio content of organic nitrogen is 1.2-2%, and the mass ratio of water content is more than 25%.
In order to ensure the rapid and stable growth of the strain MTZ102, the ratio of the culture solution in the first step is as follows: 0.15-0.25 g of yeast powder, 6-14 g of molasses and KH2PO4 0.01～0.02g，K2HPO40.01-0.02 g, 3-9 g of soybean meal, 0.5-1.5 g of wheat bran, and adding water to 1L, wherein the pH value is 6-8; after the preparation of the culture solution is finished, sterilizing at 121 ℃ for 20min
In order to ensure that the proportion of nitrogen, phosphorus and potassium in the organic fertilizer meets the growth of plants, the mass ratio of total nitrogen content in the microbial organic fertilizer in the second step is 4-5%, more than 90% of the total nitrogen content in the microbial organic fertilizer is organic nitrogen, the mass ratio of total nitrogen, phosphorus and potassium nutrients is 6-10%, and the mass ratio of organic matter content is not less than 45%.
In order to utilize the agricultural organic waste more efficiently, in the second step, the agricultural organic waste is one or more of straw, bran, bean pulp and wheat bran.
The specific fermentation medium formulation and conditions were as follows (1L): 0.2g of yeast powder, 10g of molasses and KH2PO4 0.01g，K2HPO40.01g, 5g of soybean meal and 1g of wheat bran, adding water to 1L, and keeping the pH value natural. Firstly, LB is used for culture to prepare fermentation seed liquid, the fermentation seed liquid is cultured for 2 to 3 days, and when the concentration of the strain reaches more than 108 cells/mL, the fermentation seed liquid is cultured in a proportion of 1: 10, inoculating the mixture into a culture medium for fermentation, and carrying out amplification culture for 20 days under the condition of 28-30 ℃, wherein the formation of a biological film can be observed on the surface of the fermentation liquor. The composition analysis of the membrane is mainly a mixture of fatty acids, organic acids, capsules and polysaccharides. Adding another non-membrane-producing Bacillus amyloliquefaciensFermenting under the same culture condition. The evaporation speed of the liquid surface of the fermentation membrane-producing strain is obviously lower than that of the fermentation membrane-producing strain, thereby achieving the water retention effect.
The microbial organic fertilizer is produced by using a bacterial strain MTZ102, wherein the content of the bacterial strain MTZ102 is multiplied by 108cfu/g or more, the mass ratio of total nitrogen content is 4-5%, more than 90% of total nitrogen is organic nitrogen, the mass ratio of total nitrogen, phosphorus and potassium nutrients is 6-10%, and the mass ratio of organic matter content is not less than 45%.
Fourthly, pot experiment:
first experiment of potted chrysanthemum coronarium
1.1 fermentation medium YAW (1L) with water-retaining functional bacteria of strain MTZ 102: 0.2g of yeast powder, 10g of molasses and KH2PO40.01g，K2HPO40.01g, 5g of soybean meal and 1g of wheat bran, adding water to 1L, and keeping the pH value natural.
1.2 application: the soil is uniformly mixed, the original pH value of the soil is 6.5, and the field water holding capacity is 60%. And (3) placing the activated single colony in 15ml of fresh LB seed liquid culture medium, carrying out shaking culture at 37 ℃ and 180rpm for 24h, then transferring the cultured liquid seed culture solution into a fermentation culture medium YAW according to the inoculation amount of 5%, and carrying out culture for 7d at 30 ℃ for later use. The potting experiment was performed in 4 treatments, 3 replicates. CK 1: control 1, adding 1mL of the YAW culture solution prepared above to each kg of un-inoculated soil, and normally cultivating for 20 days without watering; CK 3: comparing with 3, adding 1mL of the YAW culture solution prepared above into each kg of un-inoculated soil, normally cultivating for 20 days, and spraying appropriate amount of water every 5 days; the treatments 1-1-1 and 1-1-2 were carried out by adding 1mL of the above-prepared fermentation broth per kg of soil, and normally cultivating for 20 days without watering. The four processes are consistent with other daily management.
1.3 results:
the specific effects are shown in the following table:
1.3.1 comparison parameters of spore fermentation water-retaining bacteria for treating crowndaisy chrysanthemum
| Serial number | Average height cm of plant | Chlorophyll content mg/g | Vc content mg/g | Yield g/granule |
| CK1 | 10.5±0.5 | 1.2±0.02 | 0.18±0.02 | 40.2±0.08 |
| CK3 | 15.8±0.6 | 1.7±0.02 | 0.25±0.01 | 61.3±0.07 |
| 1-1-1 | 14.6±0.4 | 1.5±0.01 | 0.25±0.01 | 60.5±0.06 |
| 1-1-2 | 15.5±0.5 | 1.7±0.03 | 0.24±0.02 | 59.1±0.09 |
1.3.2 spore fermentation water-retaining bacteria treatment of 1-1-1 and 1-1-2 of crowndaisy chrysanthemum compared with CK1 to increase (%)
| 1-1-1 | 1-1-2 | CK3 | |
| Increase in plant height | 39.05 | 47.62 | 50.48 |
| Increased chlorophyll content | 25.00 | 41.67 | 41.67 |
| Increased Vc content | 38.89 | 33.33 | 38.89 |
| Yield improvement | 50.50 | 47.01 | 52.49 |
As can be seen from the data, the yield and average planting height of the crowndaisy chrysanthemum (1-1-1 and 1-1-2) planted by the water retention functional bacteria fermentation medium YAW produced by the strain MTZ102 are obviously higher than the content of Vc and chlorophyll which are obviously higher than the CK1 without watering; compared with CK3 watered normally, the yield, average planting height, Vc and chlorophyll content and other parameters are basically equal, which shows that the water-retaining functional bacteria fermentation culture medium YAW produced by the strain MTZ102 has good effects on increasing the quality and yield of the crowndaisy chrysanthemum under the condition of no watering.
Second) experiment of potted corn
2.1 fermentation medium YAW (1L) with water-retaining functional bacteria of strain MTZ 102: 0.2g of yeast powder, 10g of molasses and KH2PO40.01g，K2HPO40.01g, 5g of soybean meal and 1g of wheat bran, adding water to 1L, and keeping the pH value natural.
2.2 the soil is uniformly mixed, the original pH value of the soil is 6.5, and the field water holding capacity is 60%. And (3) placing the activated single colony in 15ml of fresh LB seed liquid culture medium, carrying out shaking culture at 37 ℃ and 180rpm for 24h, then transferring the cultured liquid seed culture solution into a fermentation culture medium YAW according to the inoculation amount of 5%, and carrying out culture for 7d at 30 ℃ for later use. The potting experiment was performed in 4 treatments, 3 replicates. CK 1: control 1, adding 1mL of the YAW culture solution prepared above to each kg of un-inoculated soil, and normally cultivating for 20 days without watering; CK 2: control 2, adding 1mL of the YAW culture solution prepared above to each kg of un-inoculated soil, normally cultivating for 20 days, and spraying appropriate amount of water every 5 days; treatments A1 and A2 were performed with 1mL of the fermentation broth prepared above per kg of soil, and were cultivated normally for 20 days without watering. The four processes are consistent with other daily management.
2.3 results:
2.3.1 spore fermentation water-retaining bacteria treatment corn comparison parameters
2.3.2 spore fermentation water-retaining bacteria treatment of corn A1 and A2 compared with CK1, the improvement (%)
| A1 | A2 | CK2 | |
| Increase in plant height | 43.63 | 46.75 | 50.12 |
| Increased chlorophyll a content | 45.10 | 43.14 | 47.06 |
| Increased chlorophyll b content | 50.00 | 50.00 | 56.25 |
| Dry weight increase on the ground | 44.44 | 42.22 | 42.22 |
From the above data, it can be seen that the dry weight, average plant height, chlorophyll a and chlorophyll b content of the corn (a1 and a2) planted with the water retention functional bacteria YAW produced by the strain MTZ102 is significantly higher than that of the unbwatered CK 1; compared with CK2 watered normally, the dry weight on the ground, the average plant height, the contents of chlorophyll a and chlorophyll b and other parameters are basically equal, which shows that the water retention function bacteria fermentation culture medium YAW produced by the strain MTZ102 has good effects on increasing the quality and the yield of the corn under the condition of no watering.
The above description is only a preferred embodiment of the present invention, and is not intended to limit the present invention, and any minor modifications, equivalent replacements and improvements made to the above embodiment according to the technical spirit of the present invention should be included in the protection scope of the technical solution of the present invention.
Claims (1)
1. The application of the microbial organic fertilizer of the microorganism bacteria in the cultivation of the crowndaisy chrysanthemum or the corn is characterized in that: the microorganism strain is Bacillus amyloliquefaciens (Bacillus amyloliquefaciens), the serial number of the strain is MTZ102, the preservation number is GDMCC 60099, and the preservation date is 2016, 10 and 31 days;
the content of bacterial strain MTZ102 in the microbial organic fertilizer of the microbial bacteria is 108The mass ratio of total nitrogen to total nitrogen is 4-5%, more than 90% of total nitrogen is organic nitrogen, the mass ratio of total nitrogen, phosphorus and potassium nutrients is 6-10%, and the mass ratio of organic matter content is not less than 45%;
the preparation method of the microbial organic fertilizer of the microbial bacteria comprises the following steps:
the first step, inoculating the strain MTZ102 into a culture solution of modified MRS type for fermentation production, wherein the conditions of the fermentation production are as follows: the culture temperature is 28-35 ℃, the stirring speed is 180-300 rpm, spores are formed in the middle and later stages of fermentation, and the bacteria or spore content in the fermentation liquor is more than or equal to 1 multiplied by 109cfu·ml-1；
Secondly, performing solid compost fermentation on the bacterial strain MTZ102 and the decomposed agricultural organic waste mixture, adding 100-200L of zymocyte liquid into each ton of solid organic material, turning the pile for 1 time every 7-12 days in the solid fermentation process, keeping the solid fermentation temperature not more than 80 ℃, and finishing the fermentation after 25-40 days to obtain the microbial organic fertilizer containing the zymocyte MTZ102 and the agricultural organic waste mixture;
in the second step, the agricultural organic waste is one or a combination of straws, bran coats, bean pulp and wheat bran;
the first stepThe medium culture solution comprises the following components in percentage by weight: 0.15-0.25 g of yeast powder, 6-14 g of molasses and KH2PO4 0.01～0.02g，K2HPO40.01-0.02 g, 3-9 g of soybean meal and 0.5-1.5 g of wheat bran, and adding water to 1L, wherein the pH value is 6-8; after the preparation of the culture solution, the culture solution is sterilized at 121 ℃ for 20 min.
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| CN103805204A (en) * | 2013-07-22 | 2014-05-21 | 河海大学 | Farmland soil microorganism water-holding product and preparation method thereof |
| CN104744182A (en) * | 2015-01-21 | 2015-07-01 | 河海大学 | Tobacco water retention biological organic fertilizer and preparation method thereof |
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| CN101875910A (en) * | 2010-04-20 | 2010-11-03 | 山东省食品发酵工业研究设计院 | Bacillus amyloliquefaciens for producing gamma-polyglutamic acid |
| CN103805204A (en) * | 2013-07-22 | 2014-05-21 | 河海大学 | Farmland soil microorganism water-holding product and preparation method thereof |
| CN104744182A (en) * | 2015-01-21 | 2015-07-01 | 河海大学 | Tobacco water retention biological organic fertilizer and preparation method thereof |
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Denomination of invention: A kind of microbial bacteria with the function of fermentation and water retention and its application Effective date of registration: 20220815 Granted publication date: 20210820 Pledgee: Bank of China Limited by Share Ltd. Guangzhou Liwan branch Pledgor: GUANGDONG SENDU ECOLOGICAL AGRICULTURE TECHNOLOGY CO.,LTD. Registration number: Y2022440000203 |