CN112481162B - Microbial compound bacterial agent and application thereof - Google Patents
Microbial compound bacterial agent and application thereof Download PDFInfo
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- CN112481162B CN112481162B CN202011364515.1A CN202011364515A CN112481162B CN 112481162 B CN112481162 B CN 112481162B CN 202011364515 A CN202011364515 A CN 202011364515A CN 112481162 B CN112481162 B CN 112481162B
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Abstract
The invention provides a microbial agent, which consists of the following strain fermentation broth in parts by weight: 10-50 parts of flavobacterium, 30-60 parts of azotobacter and 10-30 parts of zoogloea. The microbial compound inoculant has the following advantages: can replace pesticides to prevent and control soil-borne diseases in the growth process of the upland rice and reduce the effect of chemical residues in grains; the toughness and the strength of the base part of the stalk can be increased, and the lodging resistance of the rice can be improved; the development of the root system can be stimulated, the development of the root system is promoted, and the activity and the ground gripping capability of the root system are improved; the nitrogen can be fixed automatically, so that the continuous nitrogen is improved for the whole growth period of the rice, and the abuse of chemical fertilizers is avoided; the yield of the drought (paddy) rice can be improved; can promote early maturity of the dry rice for about two weeks, shorten the growth period and enable the rice to enter the market in advance.
Description
Technical Field
The invention belongs to the technical field of agricultural microorganisms, and particularly relates to a microbial compound inoculant capable of increasing the yield of rice and shortening the mature period of the rice and application thereof.
Background
The rice is used as an important support for agricultural food in China, and the stability of the yield and the quality of the rice directly concerns the food safety bureau in China. From the development situation of agriculture in China, the planting area of rice is 3000 ten thousand hectares per year, which accounts for about 1/3 of the total area of grain crops, and the yield accounts for about 40% of the total yield of the grain crops. China and India are the origin of rice, and the Yangtze river basin seven thousand years ago begins to plant rice. The rice planting area is very wide from the world, and the total yield is slightly lower than that of wheat. The conditions suitable for rice planting are high temperature, high humidity and short sunshine, the optimum seedling germination temperature is 28-32 ℃, the optimum ear differentiation temperature is 30 ℃ and the optimum ear emergence temperature is 25-35 ℃. In the planting management of rice, the stage from the ear differentiation to the filling stage is the key stage, and the heading stage needs to supplement a large amount of water and nutrition, increase the root activity and improve the stem and leaf functions. In order to ensure high and stable yield of rice, a rice planting management system is very important.
The rice planting mode generally comprises 7 steps: soil preparation, seedling raising, seedling transplanting, weeding, deinsectization, fertilization, irrigation, drainage and mowing. The soil preparation is to turn over soil and air-ventilate and dry; the fertilizer application mainly comprises base fertilizer and additional fertilizer, 2-3 tons of organic fertilizer can be used to be matched with the compound fertilizer, and the additional fertilizer is mainly nitrogen fertilizer in the prior art. The prior fertilizing and transplanting method is a synchronous side deep fertilizing technology for transplanting rice by a rice machine. Common diseases of rice include rice blast, bacterial leaf blight, banded sclerotial blight, rice leaf miner, mud worm and rice stem borer. And the prevention and control of rice diseases and insect pests are mainly realized through chemical protection. Imidacloprid insecticide is used for preventing and controlling rice leaf miner, and Sukebi missible oil or emamectin benzoate is used for preventing and controlling rice bollworm, deltamethrin (killing by enemy) or cyhalothrin through stem and leaf spraying.
The rice planting process needs seedling raising, rice transplanting, land preparation, soaking and water drainage, the process is labor-consuming and troublesome, and the operation is very troublesome. The current rural labor force is transferred to a city, and the labor force left in the agricultural planting industry is seriously insufficient. The complicated operation of rice planting makes old labor force unable to support, and a simple, high-efficiency, high-yield and stable-yield operation mode is very urgently needed.
The agricultural chemical is usually used by farmers for preventing and treating rice blast by kasugamycin, tricyclazole, polyoxin, carbendazim, tebuconazole and other medicaments, and the molinate-methyl or procymidone water dispersible granules are also used for stem and leaf spraying to prevent and treat rice sheath blight. The agents used to control pests cause air pollution, water pollution and a certain amount of pesticide residues. These agents enter the human body in various ways, threatening the health of humans.
In the current rice planting process, farmers often apply excessive nitrogen fertilizer to quickly plant soil nutrients, neglect to condition the physical and chemical structure of soil, even cause soil hardening and destroy the acid-base balance of the soil. The long-term application of chemical fertilizers causes soil recession, soil texture collapse, beneficial microorganisms in soil reduction, harmful plant diseases and insect pests in soil increase and the like, and the current fertilizer application mode is urgently needed to be renovated.
Easy to fall down. Due to the fact that paddy field planting is adopted, rice root systems are not developed, rice seedlings are often planted in the paddy field too shallowly, the rice root systems are not well stretched, and the ground grabbing is not firm. Too long rice internodes, vigorous surface growth and loose stalk tissue.
The nitrogen fertilizer consumption is large. When traditional rice is planted, farmers often apply excessive nitrogen fertilizer for increasing yield and stabilizing yield, wherein the nitrogen fertilizer is applied to the rice by 40 kg/mu and the phosphate fertilizer is applied to the rice by 6 kg/mu (the utilization rate is only 30-35 percent), which causes excessive growth of the rice, green and late maturity and excessively thin stalks; excessive nitrogen fertilizer causes water pollution, eutrophication of rivers and lakes and groundwater pollution.
The annual agricultural water accounts for more than 70% of the national water, and the rice water accounts for more than 70% of the total agricultural water, and is the first major water consumer. Each kilogram of rice produced needs 1-2 tons of water. The water consumption for planting the rice is 800-1000 square per mu, and mainly comprises three parts of leaf surface transpiration, water surface evaporation and underground leakage. Especially, the transpiration is vigorous from the differentiation period of young ears to ear-picking flowering, which is the period of highest physiological water demand and most concentrated for rice.
In the growth process of rice and dry rice, farmers traditionally use more nitrogen fertilizer and less phosphorus-potassium fertilizer, so that the root system is not developed, the root system cannot deeply penetrate into the field, the surface root system is limited in expansion, the activity of the root system is low, and the capacities of absorbing nutrition and transmitting substances are poor.
Disclosure of Invention
In order to solve the technical problems, the invention provides a microbial inoculum which is prepared by mixing fermentation broth of flavobacterium, azotobacter and zoogloea, and the yield of rice seeds can be improved and the maturation period of the rice seeds can be shortened by treating the microbial inoculum.
The invention is realized by the following technical scheme:
a microbial agent is composed of the following strain fermentation broth in parts by weight: 10-50 parts of flavobacterium, 30-60 parts of azotobacter and 10-30 parts of zoogloea.
Preferably, the microbial agent consists of the following strain fermentation liquor in parts by weight: 30-40 parts of flavobacterium, 30-50 parts of azotobacter and 15-30 parts of zoogloea.
Preferably, the microbial agent consists of the following strain fermentation liquor in parts by weight: 30 parts of flavobacterium, 40 parts of azotobacter and 20 parts of zoogloea.
Preferably, the flavobacterium may be an autotrophic flavobacterium (Xanthobacter), flavobacterium (Xanthobacter flavus), or Xanthobacter aminoxide (Xanthobacter aminoxidans), or flavobacterium marigold (Xanthobacter tagetidis).
More preferably, the flavobacterium is autotrophic flavobacterium (Xanthobacter autotrophicus).
Preferably, the azotobacter may be Azospirillum brasilense (Azospirillum brasilense) or Azospirillum irakense (Azospirillum irakense) or Azospirillum lipolyticum (Azospirillum lipoferum).
More preferably, the azotobacter is Azospirillum brasiliensis (Azospirillum brasilense).
Preferably, the Zoogloea may be a Zoogloea resiniphila (Zoogloea resiniphila) or Zoogloea ramigera (Zoogloea ramigera) or a Dunaliella zoogloeoides (Duganella zoogloeoides) or a zoogloeoides (Shinella zooeoides).
More preferably, the zoogloea is a zoogloea like bacterium (Shinella zooloeoides).
The invention also provides application of the microbial agent in shortening the rice maturation period.
The invention also provides application of the microbial agent in improving the yield of rice.
The invention has the beneficial effects that: the microbial compound inoculant has the following advantages: can replace pesticides to prevent and control soil-borne diseases in the growth process of the upland rice and reduce chemical residues in grains; the toughness and the strength of the base part of the stalk can be increased, and the lodging resistance of the rice can be improved; the development of the root system can be stimulated, the development of the root system is promoted, and the activity and the ground grabbing capacity of the root system are improved; the nitrogen can be fixed automatically, so that the continuous nitrogen is improved for the whole growth period of the rice, and the abuse of chemical fertilizers is avoided; can improve the yield of the drought (paddy) rice; can promote early maturity of the dry rice for about two weeks, shorten the growth period and enable the rice to enter the market in advance.
Drawings
FIG. 1 is a schematic diagram showing that the rice yield is basically stable after the composite microbial inoculum is applied to replace pesticides.
FIG. 2 is a schematic diagram of the development of rice roots in the control group and the treatment group.
Detailed Description
In order to more concisely and clearly demonstrate technical solutions, objects and advantages of the present invention, the following detailed description of the technical solutions of the present invention is provided with reference to specific embodiments and accompanying drawings. The various microorganisms in the invention can be purchased in the market or obtained from China general microbiological culture Collection center or NTCC type culture Collection center, and the preparation method of various microorganism culture solutions adopts corresponding standard culture media and culture solutions according to a conventional culture method.
Example 1 preparation of microbial Agents
1. Screening of functional strains
(1) Hydroxide bacteria
The hydrogen-oxidizing bacteria can utilize H 2 Assimilation of CO in the Presence of hydrogenase 2 And H 2 Synthesizing the substance for chemoautotrophy. In the invention, the H of the strain is determined by gas chromatography 2 The absorption capacity. Purifying the yellow bacillus: an autotrophic flavobacterium (Xanthobacter autotropicus), a flavobacterium flavum (Xanthobacter flavus), an aminoxide flavobacterium (Xanthobacter aminoxidans), or a flavobacterium marigold (Xanthobacter tagetidis); pseudomonas (Pseudomonas sp): pseudomonas hydrogenphaga (Pseudomonas hydrogenovora), rhodopseudomonas palustris (Rhodopseudomonas palustris); rhodococcus genus: inoculating Paracoccus sedimentans (Paracoccus sediminis) and Paracoccus marcusii (Paracoccus marcusii) into R2A inclined plane, sealing after bacterial coating, and using the R2A inclined plane inoculated by clear water as a control. Introducing a determined quantity H into the test tube 2 Thoroughly mixed, and then subjected to a procedure to determine H in the above-mentioned various pairs of hydrogen oxidizing bacteria 2 Absorption, in a closed tube, determined by gas chromatography, of the initial H 2 Concentration and H after three days 2 To calculate the absorption H 2 The size of the capability. The results are shown in table 1:
TABLE 1
Because the hydroxide bacteria all have the absorption of H 2 But the different species of the hydrogen oxidizing bacteria have the absorption of H 2 Are different from each other, and are screened for H uptake 2 The strain with the strongest capability is beneficial to generating the synergistic effect with other strains. Thus, in the above experiments, the inventors selected several different hydroxide bacteria and determined that they absorbed H 2 The results in Table 1 show that, in the case of the hydroxide bacteria, it is clear that H is absorbed by the xanthobacter 2 Has the strongest capacity, therefore, the yellow bacillus is selected as the microbial agent to absorb H 2 The functional bacterium of (1), wherein H is absorbed by the autotrophic yellow bacillus 2 The strain is selected as a subsequent experiment, is purchased from China general microbiological culture Collection center and has the preservation number of CGMCC 1.6351。
(2) Azotobacteria
The azotobacter activity determination method comprises the following operation steps: A15X 150mm screw glass tube was added with 5mL of modified nitrogen-fixing medium to prepare a slant, and then inoculated with Azotobacter bailii (Azotobacter beijerinckii), azotobacter asiaticum (Azotobacter armeniaca), azotobacter chroococcum (Azotobacter chroococcum), azotobacter calcoaceticus (Azotobacter chroococcum), azotobacter irasciaeli (Azotobacter iracilakense) or Azotobacter brasiliensis (Azotobacter brasiliensis) respectively and cultured at 28 ℃. The blank slant was inoculated with clear water as a negative control. After three days of culture, the rubber stopper was replaced, acetylene gas was injected to a final concentration of 10%, the mixture was sealed with a medical tape, and after three days of culture, 100. Mu.L of the reaction gas was taken, the amount of ethylene produced was measured by a gas chromatograph, and the azotase activity of the strain was calculated according to the formula. Nitrogenase activity (nmol/mg. H) = C 2 H 4 nmol/[ amount of mycoprotein (mg). Times.reaction time (h)]Wherein (C) 2 H 4 nmol＝1000×C 2 H 4 Volume (. Mu.L). Times.273 XP/[ 22.4 × (273 + t ℃ C.). Times.760]Wherein P is gas pressure (mm Hg) and t is reaction temperature).
The method for measuring the mycoprotein content is as follows: washing thallus Porphyrae on the inclined plane of the test tube into a centrifuge tube with 5mL of normal saline, collecting thallus, adding 3mL of 0.5M NaOH into the thallus, boiling for 5min in boiling water, adding 3mL of 0.5M HCl, mixing, centrifuging, taking 1.0mL of supernatant, adding 5mL of Coomassie brilliant blue solution, mixing on a vortex mixer, developing for 3 min, measuring the light absorption value A595 at 595nm, and calculating the mycoprotein content according to a bovine serum albumin standard curve. The results are shown in table 2:
TABLE 2
Because the azotobacter has the nitrogen fixing function, but the azotobacter of different types has different azotobacter capabilities, the azotobacter with the strongest azotobacter screening is beneficial to producing the synergistic effect with other strains. Therefore, in the above experiment, the inventors selected several different nitrogen-fixing bacteria and measured the nitrogen-fixing effect. The experimental result shows that the azospirillum has the strongest azozyme activity, wherein the azospirillum brasilense has the strongest nitrogen absorption capacity, the azospirillum brasilense is selected as a subsequent experiment, the strain is purchased from China general microbiological culture collection center, and the collection number of the strain is CGMCC1.10379.
2. Cultivation of the Strain
Culture of yellow bacillus: inoculating autotrophic yellow bacillus into R2A liquid culture medium, shaking and culturing for 48h at 28 deg.C to obtain bacterial suspension, and performing amplification culture until the thallus concentration is not less than 10 6 mL, secondary cultures were obtained. 1L of the medium contained yeast 0.5g, tryptone 0.25g, peptone 0.75g, glucose 0.5g, starch 0.5g, dipotassium hydrogen phosphate 0.3g, magnesium sulfate 0.024g, and sodium pyruvate 0.3g, and pH 7.2. + -. 0.2.
B, culturing azotobacter: inoculating Azospirillum brasilense into nitrogen-free liquid culture medium, culturing at 28 deg.C for 48 hr to obtain bacterial suspension, and performing amplification culture until the thallus concentration is not less than 10 6 mL, secondary cultures were obtained. 1L of the medium contained mannitol or 10g, dipotassium hydrogen phosphate 0.2g, magnesium sulfate 0.2g, sodium chloride 0.2g, calcium sulfate 0.2g, and calcium carbonate 5g, and the pH was 7.0 to 7.2.
C, culturing zoogloea: inoculating the zoogloea pseudoalternifolia into an LB liquid culture medium for culture at the temperature of 35 ℃ and 150r/min for 36-72 h by shaking culture to obtain a bacterial suspension, and then carrying out amplification culture until the concentration of thalli is not less than 10 6 mL, secondary cultures were obtained. The 1L medium contained 10g of tryptone, 5g of yeast extract, 10g of NaCl, and 7.2. + -. 0.2 pH.
Counting viable bacteria, and detecting the number of autotrophic flavobacterium, azospirillum brasilense, and Tremella mobilis of 1 × 10 6 ～1×10 10 Mixing the components per ml according to the parts by weight of fermentation liquor: 30 parts of autotrophic flavobacterium, 40 parts of azospirillum brasilense and 20 parts of zoogloea pseudozoogloea.
Example 2 Effect of the microbial Agents of the invention on Rice lodging
1. Preparation before broadcast
Variety selection: is suitable for direct seeding in dry land and is planted by selecting Longqing rice No. 2.
Land parcel selection: the land is flat, drip irrigation equipment under the film is needed in the land, and previous crops cannot have phytotoxicity.
Preparing seed fertilizer: the phosphorus and potassium supplement is emphasized, and monoammonium phosphate and potassium chloride (suitable for alkaline soil) are applied to farmyard manure in a matching way.
Soil is deeply loosened: the land is rotary-ploughed for 2 times, the soil is broken into the ground after land preparation, and the land with phytotoxicity is required to be raked in autumn, so that phytotoxicity of previous crops is reduced.
2. Seeding
Seed treatment: floating impurities such as blighted grains and the like with saline water, removing the impurities, cleaning with clean water, and drying seeds in the sun, thereby improving the germination rate and the germination potential of the seeds.
Sowing time: the ground temperature of 5 cm is stable at 10 ℃, the sowing time is suitable, and the sowing time is generally 4 months and 25 days to 5 months and 5 days per year.
Seed treatment: treatment group: the rice seeds sunned in the sun are soaked in the composite microbial inoculum prepared in the embodiment 1 for 4 hours, and are aired to be not sticky and then are sowed. Control group: and (5) directly sowing.
Sowing and film mulching: according to the variety and the sowing time, 8-10 kilograms of seeds per mu are sown by a special rice dry farming sowing machine, the film covering, the drip irrigation pipe laying and the sowing earthing are completed at one time, the sowing depth of a shallow layer machine is 1-2 centimeters, 8 (4 multiplied by 2) rows are sown every time, the row spacing is 25 centimeters (the drip irrigation pipes are laid between the large rows), the row spacing is 12 centimeters, the hole spacing is 12 centimeters, and about 15 seeds are sown in each hole. The mulching film is selected from a black film with the thickness of 0.016 mm, and the width of the film is 170 cm. Immediately drip irrigation water after sowing.
3. Field management
Root spraying: after 5-6 leaves emerge from rice seedlings and the field weeding operation is carried out for one week or 10 days, the treatment group uses the composite microbial inoculum prepared in the example 1 according to the proportion of 1 to 10 to mix well water to spray roots on the rice seedlings, and the control group uses the well water to drip irrigation.
Topdressing: in the middle stage, 6-8 times of water dripping and 3 times of water dripping topdressing compound fertilizer are needed, the dosage per mu is about 5 kilograms, 3-4 leaf nitrogen fertilizer topdressing is carried out once in the early tillering stage, 6-7 leaf topdressing is carried out once in the 6-7 leaf stage, and 2.5 kilograms of potassium chloride is added in 5 kilograms of urea topdressing per mu. Approximately half of nitrogen fertilizer is applied after dropping water in the booting stage of the treatment group, and microbial agent is applied. No microbial agent is applied in the control group during the booting stage by dropping water and topdressing nitrogen fertilizer. And in the grain filling stage of the treatment group of rice, monopotassium phosphate matched with a microbial agent is sprayed on the leaf surfaces. And potassium dihydrogen phosphate is sprayed on the control group rice in the filling stage without being sprayed on the leaf surfaces with the microbial agent.
Controlling diseases, pests and weeds: grass does not grow under the black mulching film, and the insecticide avoids the damage to microorganisms when the insecticide kills the insects. The disease prevention is carried out by spraying 1.5 kg of 30 percent Xinkewensan 100 ml or 40 percent isoprothiolane (Fushiyi) to 300 times of water at the beginning of the later seventy-months of 6 months. 750 g of 40% omethoate missible oil is used for preventing leaf miner, and 450 kg of water is sprayed.
4. Harvesting
After the dry rice is completely ripe, more than 95% of rice ears are yellow, and the rice ears are harvested by a combine harvester. The rice lodging performance was observed, and the results are shown in table 3:
TABLE 3
"+" indicates significance of treatment versus control, P < 0.05.
As can be seen from Table 3, compared with the control group, the microbial inoculum of the invention is used for soaking rice seeds before sowing, and the rice seedlings are subjected to root spraying treatment after 5-6 leaves emerge, so that the thickness of the second internode of the treatment group is obviously improved by 23%, the plant height is reduced by 8%, the gravity height is reduced by 11%, and the lodging rate is reduced by 93%. The compound microbial inoculum of the invention obviously reduces the lodging rate of the upland rice Longqing rice No. 2 and improves the lodging resistance.
Example 3 Effect of microbial Agents of the invention on Rice production
The method of example 2 was followed to plant Longqing rice No. 2, and the rice yield was observed and recorded, and the results are shown in FIG. 1, tables 4 and 5:
TABLE 4
| Group (n = 20) | Thousand Kernel weight (g) | Grain number per ear | Single ear weight (g) | Effective spike number (10) 4 hm -2 ) |
| Control group | 25.43±1.25 | 145.18±6.21 | 3.01±0.21 | 276.65±5.84 |
| Treatment group | 29.55±2.33* | 177.42±5.11* | 3.76±0.85* | 291.47±4.23* |
"+" indicates significance of treatment versus control, P < 0.05.
TABLE 5
As can be seen from Table 4, compared with the control group, the treated group increased the thousand kernel weight of rice by 8%, the number of ears by 14%, the number of single ears by 10%, and the effective number of ears by 2%. The result shows that the composite microbial inoculum of the invention obviously improves the yield of the rice Longqing rice No. 2, which is realized under the condition that the nitrogen fertilizer is reduced by half, and the composite microbial inoculum of the invention can reduce the application of the nitrogen fertilizer and has the function of biological nitrogen fixation to replace the nitrogen fertilizer. As can be seen from the table 5 in the figure 1, the composite microbial inoculum of the azospirillum chrysosporium can replace the application of pesticides by applying the composite microbial inoculum of the azospirillum chrysosporium in the dry farming rice planting mode, and the yield of rice is basically stable.
Example 4 Effect of the microbial Agents of the invention on the maturation stage of Rice
The method of example 2 was followed to plant Longqing rice No. 2, and the maturity of rice was observed and recorded, and the results are shown in Table 6:
TABLE 6
| Group (n = 20) | Average time to maturity (day) |
| Control group | 124 |
| Treatment group | 103 |
As can be seen from Table 6, the treatment group shortened the growth period required for rice maturation by 21 days on average, compared to the control group. The results of the embodiment 2 show that the composite microbial inoculum of the invention obviously improves the yield of the rice Longqing rice No. 2, but shortens the growth period, promotes the premature ripening and can harvest the rice in advance.
Example 4 Effect of the microbial Agents of the invention on Rice root development
The maturity of rice was observed and recorded by planting Longqing rice No. 2 according to the method of example 2, and the results are shown in FIG. 1. From FIG. 1, it can be seen that the treated group has developed root system, strong root system and high toughness compared to the control group. Therefore, the complex microbial inoculum can promote the development of the rice root system and obtain a more developed root system.
Example 5 Effect of the microbial Agents of the invention on soil improvement
The method of example 2 was followed to plant Longqing rice No. 2, and the rhizosphere soil of the rice in the control group and the treated group was collected at the beginning of the August, and compared with the activity of the soil within 5mm from the root system.
1. Statistics of soil microorganism types: analyzing the number of the hydroxide bacteria by using MSA culture medium; calculating the number of bacteria by using a beef extract peptone culture medium; testing the quantity of the actinomycetes by using a starch ammonium agar culture medium; the number of fungi was counted on potato-sucrose agar medium.
2. And (3) determination of dehydrogenase activity: using the TPF (triphenylformazan) colorimetric method, colorimetric quantitative analysis was performed based on the degree of chroma that produces the red color, using a hydrogen atom known to be accessible to the dehydrogenases by TTC, the acceptor species becoming TPF upon acceptance of hydrogen. Two centrifuge tubes were filled with soil samples, one added with 5mL of 5g/LTTC (triphenyltetranitrogen chloride) solution, and the other with 5mL of 0.2mol/L Tris-HCl as a reference. 0.1mol/L glucose was added to the tube (2mL) and reacted at 37 ℃ for 24 hours. 2 drops of concentrated sulfuric acid were added to terminate the reaction, 5mL of toluene was added, and the mixture was shaken for 30min. And (4) centrifuging, taking the upper layer solution at the wavelength of 485nm, taking the solution without TTC as a control, carrying out colorimetric determination on the optical density of each sample solution, and calculating the TPF content of the sample solution. And (5) drawing a standard curve, wherein the concentration of TPF is used as an abscissa, and the absorbance value is used as an ordinate. The enzyme activity was expressed as the amount of TPF produced as a reduction product of TTC [ TPF,. Mu.g/g (dry soil) ] (24 hours) -1 ]. Soil dehydrogenase Activity [ TPF, μ g/g (dry soil)]= cxv/dwt; c is the concentration of TPF in the filtrate (μ gm/L); v is filtrate volume (mL); and dwt is the dried soil weight (g).
3. Determination of catalase activity: the activity of catalase was expressed by the amount of hydrogen peroxide remaining from the hydrogen peroxide decomposition reaction by potassium permanganate titration. Soil catalase activity [ KMnO 4 mL/g (dry soil)]= (V0-V)/dwt; v0 and V are the number of milliliters (mL) of blank titration and sample titration of 0.2mol/L potassium permanganate; and dwt is the dried soil weight (g).
4. Determination of urease activity: indophenol blue colorimetry is adopted. Leached NH 4+ Reacting hypochlorous acid and phenol in a strong alkaline medium to generate water-soluble indophenol blue, NH 4+ Proportional to the shade of the color. Soil urease Activity [ NH ] 4+ Mg/g (dry soil) (24 h) -1 ]= (C × 10)/dwt; c is the enzyme activity value (NH) in the soil filtrate 4+ mg/mL); 10 is the dilution multiple of the soil solution; and dwt is the dried soil weight (g).
5. Detection of invertase Activity: the 3, 5-dinitrosalicylic acid colorimetric method is adopted. Invertase can hydrolyze sucrose, which is a non-reducing sugar, into glucose and fructose, and the amount of reducing sugar produced is determined to indicate the activity of the invertase. The activity of the invertase was expressed in ml of glucose per unit soil weight.
The detection results are shown in tables 7 and 8:
table 7: statistical results of soil microorganism classes
Table 8: content detection results of four enzymes
As can be seen from tables 7 and 8, the number of the treated group of the bacteria with hydrogen peroxide was significantly increased as compared to the control group, and similarly, the number of the treated group of the bacteria, actinomycetes and fungi was increased to various degrees. The microbial quantity of the rice rhizosphere soil after the composite microbial inoculum is applied is increased. The contents of four enzymes, namely dehydrogenase, catalase, urease and invertase, in the soil of the treatment group are increased, which shows that the soil microbial population and the activity of the rice rhizosphere soil after the composite microbial inoculum is applied are increased, the decomposition and conversion rate of carbohydrates in the soil is increased, hydrogen peroxide generated by biochemical reaction in the soil is reduced, the hydrolysis of amide peptide bonds of nitrogen-containing organic compounds urea molecules in the soil is increased (the generated plant nitrogen ammonia is increased), and the soil is improved.
Example 6
The embodiment provides a microbial agent, which comprises the following components in parts by weight: 10 parts of autotrophic flavobacterium, 30 parts of azospirillum brazilian and 30 parts of zoogloea sp.
Example 7
The embodiment provides a microbial agent, which comprises the following components in parts by weight: 50 parts of autotrophic flavobacterium, 60 parts of azospirillum brasilense and 10 parts of zoogloea pseudostellata.
Example 8
The embodiment provides a microbial agent, which comprises the following components in parts by weight: 40 parts of autotrophic flavobacterium, 50 parts of Azospirillum brasilense and 15 parts of Tremella mobilis.
Example 9
The present example provides a microbial agent, and the only difference between the present example and example 1 is that flavobacterium flavum is used instead of autotrophic flavobacterium flavum.
Example 10
The present example provides a microbial agent, and the only difference between the present example and example 1 is that the amine oxide yellow bacillus is used to replace the autotrophic yellow bacillus.
Example 11
The present example provides a microbial agent, and the only difference between the present example and example 1 is that xanthobacter marigold is used instead of autotrophic xanthobacter.
Example 12
The present example provides a microbial agent, and the only difference between the present example and example 1 is that azospirillum irascitarum is used instead of azospirillum brasilense.
Example 13
The present example provides a microbial agent, and the only difference between the present example and example 1 is that azospirillum lipolyticum is used instead of azospirillum brasilense.
Example 14
The present example provides a microbial agent, and the only difference between the present example and example 1 is that zoogloea resinophila is used instead of zoogloea.
Example 15
The present example provides a microbial agent, and the only difference between the present example and example 1 is that zoogloea ramigera is used instead of zoogloea sp.
Example 16
This example provides a microbial agent, and the only difference between this example and example 1 is that the tunica donii is used instead of the tunica pseudotunica.
Comparative example 1
The only difference between comparative example 1 and example 1 is that the microbial agent of comparative example 1 does not contain xanthobacter. The total parts by weight are in accordance with the examples.
Comparative example 2
The only difference between comparative example 1 and example 1 is that the microbial agent of comparative example 1 does not contain azotobacter. The total parts by weight are in accordance with the examples.
Comparative example 3
The only difference between comparative example 1 and example 1 is that the microbial agent of comparative example 1 does not contain zoogloea sp. The total parts by weight are in accordance with the examples.
Comparative example 4
The only difference between comparative example 4 and example 1 is that the microbial agent of comparative example 4 contains only xanthobacter xanthus and azotobacter. The total parts by weight are in accordance with the examples.
Comparative example 5
The only difference between comparative example 5 and example 1 is that the microbial agent of comparative example 5 contains only azotobacter and zoogloea. The total parts by weight are in accordance with the examples.
Comparative example 6
The only difference between comparative example 6 and example 1 is that the microbial agent of comparative example 6 contains only flavobacterium and zoogloea. The total parts by weight are in accordance with the examples.
The microbial agents of examples 6 to 16 and comparative examples 1 to 6 were treated with rice seeds and roots, and the effects on the rice maturity stage were compared (using the method of example 2), with the results shown in table 9:
TABLE 9
As is clear from the above table, the maturation period of the rice plants treated with the microbial agents of examples 1 and 6 to 16 was significantly shortened. This is because the microbial agents of examples 1 and 7 to 16 contain flavobacterium, azotobacter and zoogloea simultaneously, while the microbial agents of comparative examples 1 to 8 lack one or more of them, so that the microbial agents of the present invention have synergistic effect of flavobacterium, azotobacter and zoogloea, and can improve yield and shorten maturation period of rice, so that rice can be listed in advance.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is specific and detailed, but not to be understood as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (3)
1. The application of the microbial agent in shortening the mature period of rice or improving the yield of rice is characterized in that the microbial agent consists of the following strain fermentation liquid in parts by weight: 10-50 parts of flavobacterium, 30-60 parts of azotobacter and 10-30 parts of zoogloea;
the flavobacterium is an autotrophic flavobacterium (Xanthobacter autotrophicus); the azotobacter is Azospirillum brasiliensis (Azospirillum brasilense); the zoogloea sp is a zoogloea sp (Shinella zooeoides).
2. The use of the microbial inoculant according to claim 1 for shortening the maturation period or increasing the yield of rice, wherein the microbial inoculant comprises the following fermentation broths of the species in parts by weight: 30-40 parts of flavobacterium, 30-50 parts of azotobacter and 15-30 parts of zoogloea.
3. The use of the microbial inoculant according to claim 2 for shortening the maturation period or increasing the yield of rice, wherein the microbial inoculant consists of the following fermentation broths of the species in parts by weight: 30 parts of flavobacterium, 40 parts of azotobacter and 20 parts of zoogloea.
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| CN113322198B (en) * | 2021-05-06 | 2023-04-07 | 广东丽豪生物农业有限公司 | Microbial agent based on xanthobacter oxamine and xanthobacter oryzae as well as preparation method and application of microbial agent |
| CN113337421A (en) * | 2021-05-06 | 2021-09-03 | 广东丽豪生物农业有限公司 | Compound microbial agent and application thereof |
| CN113337420A (en) * | 2021-05-06 | 2021-09-03 | 广东丽豪生物农业有限公司 | Composite nitrogen-fixing microbial agent and preparation method and application thereof |
| CN115067362B (en) * | 2022-08-15 | 2023-08-11 | 湖南省植物保护研究所 | Sterilization composition containing rhodopseudomonas palustris and tricyclazole |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101524089A (en) * | 2009-04-17 | 2009-09-09 | 上海创博生物技术有限公司 | Compound microbial preparation for promoting paddy growth and preparation method thereof |
| CN103834565A (en) * | 2012-11-27 | 2014-06-04 | 南阳奇伟微生态基因科技开发有限公司 | Micro-ecological flora of crop root system probiotics and application thereof |
| CN106810370A (en) * | 2017-02-22 | 2017-06-09 | 广州聚禅现代农业研究院有限公司 | A kind of microbial manure containing chelated microelements and preparation method thereof |
| CN108486001A (en) * | 2018-03-13 | 2018-09-04 | 常州大学 | A kind of photosynthetic microbial inoculum of new and effective active selenium-rich and its preparation method and application for paddy growth |
| CN110241041A (en) * | 2019-05-31 | 2019-09-17 | 南京工业大学 | A kind of complex microorganism preparations, preparation method and application |
-
2020
- 2020-11-27 CN CN202011364515.1A patent/CN112481162B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101524089A (en) * | 2009-04-17 | 2009-09-09 | 上海创博生物技术有限公司 | Compound microbial preparation for promoting paddy growth and preparation method thereof |
| CN103834565A (en) * | 2012-11-27 | 2014-06-04 | 南阳奇伟微生态基因科技开发有限公司 | Micro-ecological flora of crop root system probiotics and application thereof |
| CN106810370A (en) * | 2017-02-22 | 2017-06-09 | 广州聚禅现代农业研究院有限公司 | A kind of microbial manure containing chelated microelements and preparation method thereof |
| CN108486001A (en) * | 2018-03-13 | 2018-09-04 | 常州大学 | A kind of photosynthetic microbial inoculum of new and effective active selenium-rich and its preparation method and application for paddy growth |
| CN110241041A (en) * | 2019-05-31 | 2019-09-17 | 南京工业大学 | A kind of complex microorganism preparations, preparation method and application |
Non-Patent Citations (4)
| Title |
|---|
| 兰州地区盐碱地小麦根际联合固氮菌分离及部分特性研究;姚拓等;《土壤学报》;20040630(第03期);全文 * |
| 复合生防菌群对连作大豆根际土壤可培养微生物区系的影响;范文艳等;《中国油料作物学报》;20120615(第03期);全文 * |
| 微生态制剂SC27对作物生长和土壤环境的影响;周培等;《上海交通大学学报(农业科学版)》;20021230(第04期);全文 * |
| 野生稻内生固氮菌多样性研究进展;刘丽辉等;《生物技术进展》;20171125(第06期);全文 * |
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