CN1475467A - Bamboo forest biological fertilizer - Google Patents
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Abstract
A biological fertilizer for bamboo forest is prepared from Paenibacillus polymyxa WG-7 and/or Bacillus licheniformis WG-8, and matrix.
Description
Technical field
The present invention relates to contain the fertilizer of microorganism, particularly the bamboo forest biological fertilizer material.
Background technology
Chemical fertilizer is easy to use, and fertilizer efficiency is fast, and the essential element of plant growth can be provided, and has played keying action to solving human problem of food and clothing.But chemical fertilizer also has many drawbacks, as: consume a large amount of energy during production, the production cost height; Can produce pollution to water and atmosphere in production and the use, the turnover rate height, the plant utilization rate is low; The excessive nitrite of using plant interior accumulation behind the chemical nitrogen fertilizer is harmful to HUMAN HEALTH; Because the chemical fertilizer element is single, is difficult to rational proportion when arranging in pairs or groups mutually, agricultural-food nutritive value and local flavor are reduced; Use chemical fertilizer, correspondingly reduce the fertilizer use and bring soil with organic matter content to reduce degradation problem under soil compaction, the soil fertility thereupon.Therefore, people attempt to seek other source of manure always.
Since D bereiner (1976) has reported that containing the firm spirillum of fat and maize root system has associative nitrogen fixation, caused international generally attention, countries in the world are to many gramineae farm crops and herbage carries out nitrogen fixing capacity mensuration and vinelandii separate, confirm that grass generally has associative nitrogen fixation, for hope has been found in the nitrogen supply that utilizes biological nitrogen fixation to solve gramineous crop.Just this is studied in the world at present, developed the association nitrogen fixation bacterial manure of many suitable gramineae farm crops from the genetically engineered improvement of bacterial strain screening, microbial inoculum application, bacterial strain and the numerous areas such as microbial ecology of root system of plant.
Present research thinks that association nitrogen fixation is between a kind of fixed nitrogen form between growing nitrogen-fixing and symbiotic nitrogen fixation, association nitrogen fixation and there is no clear and definite boundary between growing nitrogen-fixing; Combination azotobacter is that a class is grown surely the vinelandii that do not form specialization structure than substantial connection on roots of plants are arranged between plant rhizosphere and plant; At a large amount of organic carbon source materials of plant rhizosphere root system excretory, for growth, breeding and the fixed nitrogen of combination azotobacter provides the carbon source and the energy, and combination azotobacter fixed nitrogen is easy to again by plant utilization, is the mutual reciprocity and mutual benefit relation between combination azotobacter and the plant; Combination azotobacter has certain host specificity and homology according to the study, nitrogen-fixing efficiency is very high with its host plant associating the time, and the host is also very high to the utilization ratio of its fixed nitrogen, though and combination azotobacter also can fixed nitrogen under spontaneous condition, but nitrogen-fixing efficiency is very big with changes in environmental conditions, as under temperature and humidity condition suitable on the artificial medium, the nitrogen-fixing efficiency of combination azotobacter is also very high, but the nitrogen-fixing efficiency of combination azotobacter is in general just lower in natural soils.
Bamboo plant also belongs to Gramineae, is the important forest plants of China torrid zone, subtropical zone, also is the especially important forest reserves of Southeast Asian countries of world many countries.Because its special growth vigor and important economic value, and more and more be subjected to the attention of countries in the world, but its Biological Nitrogen Fixation Researches is not appeared in the newspapers.Applicant proposed bamboo plant should have the association nitrogen fixation characteristic as the major member of grass viewpoint in the world first in 1994.Through research for many years, not only confirm the same certain associative nitrogen fixation that has of bamboo plant with other gramineae farm crop, and be separated to a large amount of combination azotobacters from multiple bamboo plant rhizosphere, after these combination azotobacters are further studied, screen the bacterial strain that the higher tool research and utilization of a few strain nitrogenase activities is worth, with them to the mao bamboon seedling, after sinocalamus latiflorus tissue cultured seedling and multiple bamboo forest in use for bamboo shoot are inoculated and are used, the proof tool obviously improves bamboo root system and bamboo grove soil nitrogen fixing capacity, increase the bamboo leaves nitrogen content, promote the bamboo especially root growth of growing, improve effects such as bamboo shoot output and plant percent.
After China solved the basic living problems substantially, environment and food-safety problem earned widespread respect at present, complied with international trend, and China is more and more higher to the demand of pollution-free food, green food, Organic food.Pollution-free bamboo shoots, green bamboo shoots and organic bamboo shoots of various places development at present, be badly in need of efficient biofertilizer, fertilizer and biological organic complex fertilizer etc., with fertilizer for replacing chemical partially or completely, simultaneously use bio-fertilizer or the alternative chemical fertilizer of fertilizer in bamboo grove, also can reduce water source and environmental pollution at the big area material.Therefore, developed with bamboo rhizosphere combination azotobacter as the major function bacterial classification, and the bamboo forest biological fertilizer that cooperates other measure to produce.Bamboo forest biological fertilizer has not only comprised the combination azotobacter from bamboo plant rhizosphere separation screening, and can be with the microorganism of other kind and/or various organic matter (fertilizer) and/or inorganic fertilizer and/or other composition compound or be mixed, form the bamboo forest biological fertilizer that a class can satisfy different bamboo plants, different production purpose, different mass requirement.
The application of associating azotobacteria fertilizer generally is by modes such as seed pelleting or microbial inoculum seed dressings on gramineae farm crop, and seed is contacted closely with vinelandii, and combination azotobacter at first occupied the root system vantage point when seed germination took root, and forms advantage at rhizosphere.Because bamboo is a perennial plant, and bamboo is seldom used planting seed, add the complicacy of bamboo plant root system underground structure, bring difficulty for using of bamboo association nitrogen fixation bacterial manure, combination azotobacter is difficult to directly apply Gen Biao and indigenous bacterium contention vantage point forms dominant microflora, after entering soil, the vinelandii that touch root system just might and indigenous bacterium competition and carry out association nitrogen fixation, and if the combination azotobacter that does not reach root system will be in dormant state or very fast death when lacking the supply of carbon source material and other nutrient.Propose according to this situation contriver, when bamboo grove is used association nitrogen fixation bacterial manure, at first to manage to guarantee the combination azotobacter abundant survival under the spontaneous condition and vigorous breeding in soil, just can make vinelandii permeate and finally to occupy the rhizosphere advantageous position to whole root system at leisure on this basis and carry out association nitrogen fixation.Preferably cooperate the necessary middle trace element of organic carbon source material and phosphorus potassium element and various plants and microorganism when therefore the bamboo grove combination azotobacter applies fertilizer simultaneously, wherein the organic carbon source material is a key factor, so-called organic carbon source material comprises rich cellulose, the organic matter of hemicellulose and various carbohydrates or fertilizer, these materials can be vinelandii abundant low molecule carbon sources such as glucose are provided after cellulose-decomposing bacterium is decomposed in soil, because the supply of competent organic carbon source and mineral element is arranged, and the water-retentivity of soil is strengthened, make combination azotobacter enter growth and breeding and the fixed nitrogen under spontaneous condition that to conform earlier behind the soil, form dominant microflora and the indigenous bacterium contention rhizosphere position that is at war with at the bamboo rhizosphere the most at leisure and carry out association nitrogen fixation.This bamboo forest biological fertilizer not only has associative nitrogen fixation, and at first also has free living nitrogen fixation.Also can cooperate various soil beneficial microorganisms such as phosphate solubilizing bacteria, potassium solubilizing bacteria in the bamboo forest biological fertilizer simultaneously, improve plant and vinelandii utilization ratio phosphorus potassium element and organic carbon source material.
Summary of the invention:
Technical problem to be solved by this invention is: provide nitrogen by biological nitrogen fixation for bamboo, when cooperating other composition such as phosphorus bacteria, potassium bacterium and fertilizer and inorganic fertilizer, also have concurrently and improve phosphorus potassium content and utilization ratio in fertilizer and the soil, further improve the soil, improve the effects such as micro-ecological environment of microorganism.Thereby using of minimizing or alternative bamboo plant chemical nitrogen fertilizer improved the bamboo shoots quality, to the demand of bio-feritlizer, avoids bamboo grove general with the pollution of chemical fertilizer to environment when adapting to nuisanceless green bamboo shoots and organic production of bamboo shoots.
Its special character of technical scheme that bamboo forest biological fertilizer of the present invention adopts is to be made of bamboo plant rhizosphere combination azotobacter and matrix, and described bamboo plant rhizosphere combination azotobacter is from fixedly Paenibacillus polymyxa (Paenibacillus polymyxa) WG-7 bacterial strain and any bacterial strain in Bacillus licheniformis (Bacilluslicheniformis) the WG-8 bacterial strain or the combination of two bacterial strains of molecular nitrogen the atmosphere of the isolating energy of bamboo plant rhizosphere.
Described from the isolating Paenibacillus polymyxa of bamboo plant rhizosphere (Paenibacilluspolymyxa) WG-7 bacterial strain and Bacillus licheniformis (Bacillus licheniformis) WG-8 bacterial strain, in on July 12nd, 2002 in China Committee for Culture Collection of Microorganisms's common micro-organisms center preservation, this centre address is: China, Beijing, the Zhong Guan-cun, deposit number is: CGMCC No.0768[Paenibacillus polymyxa (Paenibacillus polymyxa) WG-7 bacterial strain]; CGMCC No.0769[Bacillus licheniformis (Bacilluslicheniformis) WG-8 bacterial strain].
Bamboo forest biological fertilizer of the present invention, can also comprise phosphorus bacteria bacillus megaterium (Bacillus megaterium, culture presevation number is: ACCC 10010) and/or potassium bacterium bacillusmusilaginosiengineering (Bacillus mucilaginosus, culture presevation number is: ACCC10013), can obtain from China Committee for Culture Collection of Microorganisms agricultural microorganism center.
Described matrix is a kind of of the existence of described bacterial classification or the liquid medium of cultivating, sorbent material, organic materials, inorganic material or several;
Described bio-fertilizer is: a) liquid bio fertilizer, b) solid fungicide, c) biological organic fertilizer and d) biologic organic, inorganic composite fertilizer any, wherein:
Liquid bio fertilizer is the liquid bio fertilizer that described bacterial classification forms after the liquid nutrient medium fermentation, and wherein the liquid medium of bacterial classification existence or cultivation comprises water, opsonigenous substances and part nutritive substance;
Solid fungicide is the solid fungicide that described liquid bio fertilizer forms after adsorbents adsorb; Or the freeze-drying microbial inoculum that after freeze-drying, forms; Or the dry microbial inoculum of spray-dried back formation.
Biological organic fertilizer is the biological organic fertilizer of described liquid bio fertilizer or solid fungicide and described organic materials formation;
Biologic organic, inorganic composite fertilizer is the biological organic and inorganic fertilizer that described liquid bio fertilizer or solid fungicide and described organic materials, inorganic material constitute;
Described freeze-drying microbial inoculum can carry out with present known various freeze-drier and the methods that are used for bacterium;
Described spraying drying microbial inoculum can carry out with present known various spray drying device and the methods that are used for genus bacillus;
Described sorbent material is a material: the mixture of one or more materials in the matter is planted in the peat composed of rotten mosses, peat, vermiculite, light calcium carbonate, kaolin, pond sludge, wilkinite, corruption;
Described organic materials is: the mixture of one or more materials in the materials such as (bagasses in vinasse, mushroom slag, sugar refinery etc.) of the organic waste in crop material, wood chip, animal excrement, municipal wastes, mud, the industrial and agricultural production; Or the organic fertilizer that behind fermentation maturity, forms again of above-mentioned organic materials;
Described inorganic material is various macroelement and the middle micro-chemical fertilizer of selling on the market etc. of containing.
The separation method of described bamboo plant rhizosphere combination azotobacter is: the bamboo root is connected soil and dug along the bamboo pocket, shake off surface dust after taking back the aseptic experiment chamber, wash down earth with tap water again; Select the healthy and strong root of living and be cut into root segment,, use 70% alcohol-pickled 5min then, remove the root segment excessive moisture with inhaling with aseptic filter paper behind the aseptic water washing 6 times again with aseptic water washing 6 times less than 1cm; Take by weighing the bright root segment of 1g respectively and put into the penicillin bottle that the D bereiner that the 2ml modification is housed does not have the nitrogen semisolid medium, empty lid bottle cap (blow-by) was cultivated 24 hours for 30 ℃ in the sterile culture case; Build bottle cap (rubber bottle cap),, inject 0.7ml acetylene, continue to cultivate after one hour, measure acetyiene reduction activity with the mouth of sealing with wax; Therefrom select activity and active higher bottled sample, do not have line separation on the nitrogen solid plate, picking list bacterium colony and repeatedly purifying then at the D bereiner that revises with inoculating needle picking nutrient solution; Bacterium behind the purifying is inserted the D bereiner that modification is housed again and is not had in the penicillin bottle of nitrogen semisolid medium, measures acetyiene reduction activity (nitrogenase activity); The bacterium of the higher acetyiene reduction activity of tool is further studied and identifies.
The isolating modified D bereiner of described bamboo rhizosphere combination azotobacter does not have nitrogen solid and semisolid medium, and its prescription is: take by weighing glucose 5.0g, sucrose 5.0g, oxysuccinic acid 5.0g, KH
2PO
40.4g, K
2HPO
4.3H
2O 0.1g, MgSO
4.7H
2O 0.2g, NaCl 0.1g, CaCl
2.2H
2O 0.02g, FeCl
3.3H
2O 0.01g, Na
2MoO
4.2H
2O0.002g puts into the beaker water dissolution successively, adds KOH 4g again, stirs to dissolve the back with pH test paper mensuration pH value of solution, regulates about its pH to 7.2 with the KOH dilute solution, adds yeast extract 0.4g again, stirs, and dissolving is settled to 1 liter.Add 2% agar during the configuration solid medium, add 0.2% agar during the preparation semisolid medium, the sterilization back is standby.
Describedly be from isolating two its grown forms of strain combination azotobacter of bamboo plant rhizosphere and physiological and biochemical property:
The essential characteristic of WG-7 bacterial strain [Paenibacillus polymyxa (Paenibacillus polymyxa)] is: cell is Gram-positive, shaft-like, size is 0.7-0.8 μ m * 3.0-3.5 μ m, and the tool end is given birth to inferior end and given birth to oval gemma, sporangiocyst expands, no PHB particle in the born of the same parents; Bacterium colony is little, and is smooth, point-like, projection, cheese look 1-2mm; The V-P reacting positive; Glucose, wood sugar, pectinose produce acid and aerogenesis; Tool catalase and amylase; Oxydase and lecithinase feminine gender, but energy decomposing casein; Nitrate can revert to nitrite; The pH5.7 growth or not among the 7%NaCl; Anti-N,O-Diacetylmuramidase; Otan (glycerine) positive; Can on anaerobic culture medium, grow; On nitrogen-free agar, grow energy fixed member attitude nitrogen; Name and be Paenibacillus polymyxa (Paenibacillus polymyxa).
The essential characteristic of WG-8 bacterial strain [Bacillus licheniformis (Bacillus licheniformis Chester)] is: cell is Gram-positive, and is shaft-like, and size is 0.8 μ m * 2.2-3.0 μ m; Give birth to oval gemma in the tool, sporangiocyst expands slightly to not expanding; No PHB particle in the born of the same parents; In pH5.7,7%NaCl and N,O-Diacetylmuramidase, all can grow; Litmus milk peptonizes, and does not produce acid; The V-P reacting positive; Catalase, oxydase and amylase are all positive; The Phospholipid hydrolase of not laying eggs, and can decomposing casein; Glucose, wood sugar, pectinose and N.F,USP MANNITOL are all produced acid; Features such as otan (glycerine) feminine gender; Can on anaerobic culture medium, grow; On nitrogen-free agar, grow energy fixed member attitude nitrogen; Name and be Bacillus licheniformis (Bacilluslicheniformis Chester).
Bamboo forest biological fertilizer of the present invention, its manufacture method is mainly:
A) liquid bio fertilizer----
1. fermentation culture Paenibacillus polymyxa (Paenibacillus polymyxa mace) WG-7 bacterial strain is made Paenibacillus polymyxa WG-7 strain liquid bio-fertilizer.
2. fermentation culture Bacillus licheniformis (Bacillus licheniformis Chester) WG-8 bacterial strain is made Bacillus licheniformis WG-8 strain liquid bio-fertilizer.
3. will be 1. and 2. two kinds of liquid bio fertilizer volume ratio on demand be mixed, just become conglomeration vinelandii liquid bio fertilizer.
4. fermentation culture bacillus megaterium ACCC 10010 bacterial strains are made bacillus megaterium ACCC 10010 strain liquid bio-fertilizers.
5. fermentation culture bacillusmusilaginosiengineering ACCC 10013 bacterial strains are made bacillusmusilaginosiengineering ACCC 10013 strain liquid bio-fertilizers.
6. with liquid bio fertilizer 4. with aforesaid liquid bio fertilizer 1. or 2. or 3. on demand volume ratio be mixed, just become the various liquid bio fertilizer that contain combination azotobacter and phosphorus bacteria.
7. with liquid bio fertilizer 5. with aforesaid liquid bio fertilizer 1. or 2. or 3. or 6. on demand volume ratio be mixed, just become the various liquid bio fertilizer that contain combination azotobacter, phosphorus bacteria and/or potassium bacterium.
The fermentation condition of WG-7 and WG-8 bacterial strain can use the general fermentation condition of known vinelandii, and optimized conditions is: inoculum size 10%, culture temperature 30-35 ℃, pH6.8-7.2,200 rev/mins of stirring velocitys, incubation time 20～30 hours, air flow 0.5～1: 1 cubic metre/1000 liters.
The test tube solid slant culture base of WG-7 and WG-8 bacterial strain can adopt general vinelandii to use substratum always, and the culture medium prescription of optimization is: glucose 0.5%, sucrose 0.5%, KH
2PO
40.04%, K
2HPO
4.3H
2O 0.01%, NH
4(SO
4)
20.1%, MgSO
4.7H
2O 0.02%, and NaCl 0.01%, CaCl
2.2H
2O 0.002%, FeCl
3.3H
2O0.001%, Na
2MoO
4.2H
2O 0.0005%, K
2SO
40.15%, yeast extract 0.06%, agar 2%, pH6.8-7.2.
Multiple substratum all can be used for the shaking table of WG-7 and WG-8 bacterial strain to be cultivated and fermentative production, and the culture medium prescription of optimization is: glucose 0.5%, sucrose 0.5%, Semen Maydis powder 1%, K
2SO
40.15%, KH
2PO
40.04%, K
2HPO
4.3H
2O 0.01%, NH
4(SO
4)
20.1%, MgSO
4.7H
2O 0.02%, and NaCl 0.01%, CaCl
2.2H
2O 0.002%, FeCl
3.3H
2O0.001%, Na
2MoO
4.2H
2O 0.0005%, peptone 0.1%, yeast extract 0.08%.
Cultivation of phosphorus bacteria bacillus megaterium and fermentation condition carry out (Chen Tingwei etc., " phosphorus bacteria ", agriculture press, nineteen fifty-nine, Beijing) by known method.The a large amount of fermention mediums of cultivating of phosphorus bacteria can be used known prescription: starch 2%, soybean cake powder 2%, ammonium sulfate 0.2%, potassium primary phosphate 1%, yeast powder 0.1%, lime carbonate 0.01%, peptone 0.1%, pH7.2-7.4.Other fermentation condition is identical with the bamboo plant combination azotobacter.
Cultivation and the fermentation condition of potassium bacterium frozen glue sample genus bacillus carry out (Chen Tingwei etc., " phosphorus bacteria ", agriculture press, nineteen fifty-nine, Beijing) by known method.The a large amount of fermention mediums of cultivating of potassium bacterium can be used known prescription: starch 10%, sucrose 0.1%, dipotassium hydrogen phosphate 0.2%, ammonium sulfate 0.1%, sal epsom 0.05%, iron trichloride 0.002%, lime carbonate 0.05%, yeast powder 0.1%, soybean cake powder 5%, pH7.2-7.4.Other fermentation condition is identical with the bamboo plant combination azotobacter.
Concrete cultivation of above-mentioned bacterium and fermentation process can carry out according to a conventional method, the method of optimizing is: adopt common microorganism purification and rejuvenation technology, respectively to each bacterial strain of above-mentioned three major types bacterium processing of purificating and rejuvenating, bacterial strain after will purificating and rejuvenating respectively again moves on the fresh solid slant tube substratum of receiving the three major types bacterium, cultivated respectively 24 hours at 30-32 ℃, treat that surface growth enters the shaking table cultivation stage after full.With above-mentioned shaking table substratum and above-mentioned fermentation condition to each bacterial strain in triangular flask on the shaking table 30-32 ℃ cultivated 20-30 hour, inoculum size with 10% when bacterial count reaches 2,000,000,000/milliliter inserts first class seed pot, when treating that the bacterium number reaches 18-20 hundred million/milliliter, inoculum size with 10% inserts the secondary seed jar, insert three grades of seeding tanks again, can improve temperature in late stage of culture, impel the bacterial growth gemma, microscopy bacterium number reaches 18-20 hundred million/milliliter, when reaching 80-100%, gemma puts jar, become various liquid bio fertilizer, they are mixed in required ratio just becomes various hybrid bacterial strain liquid bio fertilizer.
B) solid fungicide----
After aforesaid sorbent material pulverizing, sterilization, add needed more than one state a) in 1. or 2. or 3. or 6. or a certain liquid bio fertilizer of 7. method preparation mix and stir, make the quantity and the water content of function yeast be controlled at the scope that fertilizer requires, just form various bamboo forest biological fertilizer solid fungicides.
The pulverizing of sorbent material and sterilization can be adopted various known method, the most well 80 mesh sieves of the material after the pulverizing, and the assorted bacterium numerical control of the material after the sterilization there is no pathogenic bacterium built in below 5%.Can adopt stirring-type high moisture material cylinder oven dry unit to high moisture material, control gas flow temperature at 600-750 ℃, 15-25 minute drying and sterilizing time; Can adopt 200 ℃ of hot-air seasoning sterilizations of hot blast cylinder dryer to common material; Sterilization back material cool to room temperature.
The ratio of sorbent material and liquid bio fertilizer does not have strict restriction, can experimentize according to the raw material condition of use and adjust.But the quality standard of described solid fungicide preferably can be optimized to: when not containing phosphorus bacteria and potassium bacterium, and the living bacteria count of combination azotobacter 〉=500,000,000/gram; When containing phosphorus bacteria and/or potassium bacterium, total function yeast living bacteria count is best 〉=600,000,000/gram, and the living bacteria count of every kind of function yeast is no less than 200,000,000/gram, and water content is controlled at below 30%, can carry out cryodrying when water content is too high and handle.
Solid fungicide is can also be at present known various to be used for the freeze drying process of bacterium and equipment and to carry out freeze-drying and handle and make the freeze-drying microbial inoculum; Or carry out drying treatment with at present known various spray drying process that is used for genus bacillus and equipment and make dry microbial inoculum.
C) biological organic fertilizer----
After the aforesaid fertilizer pulverizing that ferments, sterilization, adding needed more than one states a) or b) a certain liquid bio fertilizer or the solid fungicide of method preparation, mix, make the quantity and the water content of various function yeast be controlled at the scope that fertilizer requires, can handle through cryodrying if moisture content is too high, make biological organic fertilizer, the biological organic fertilizer of making also further granulation make the particulate state biological organic fertilizer.
D) biologic organic, inorganic composite fertilizer----
With the aforesaid fertilizer that ferments pulverize, after the sterilization, add an amount of inorganic fertilizer as required through pulverizing, mix thoroughly, adding needed more than one again states a) or b) the liquid bio fertilizer or the solid fungicide of method preparation, mix, make the quantity and the water content of various function yeast be controlled at the scope that fertilizer requires, can handle through cryodrying, make biologic organic, inorganic composite fertilizer if moisture content is too high.
Also can be with composite organic-inorganic fertilizer, biological organic fertilizer granulation respectively, remix forms biologic organic, inorganic composite fertilizer when using.
The pulverizing of organic materials and sterilization can be adopted the whole bag of tricks, and the assorted bacterium numerical control of the material after the sterilization there is no pathogenic bacterium built in below 5%.
Described biological organic fertilizer or biologic organic, inorganic composite fertilizer can also be by following method productions: aforementioned unleavened organic materials is pulverized, inserting needed more than one states a) or b) the liquid bio fertilizer or the solid fungicide of method preparation, adopt known various fermentation process and equipment at present, with combination azotobacter or combination azotobacter, phosphorus bacteria and/or potassium bacterium and organic raw material co-fermentation, form biological organic fertilizer; Add an amount of inorganic fertilizer as in the fermentation or fermentation back, just form biologic organic, inorganic composite fertilizer.Because the bacterial classification of described production bamboo forest biological fertilizer is bacillus, bacterial classification can form gemma and enter dormant state when leavening temperature is high, and bacterial classification can be bred rapidly when leavening temperature is suitable.
The add-on of organic materials and inorganic material can be adjusted according to the quality requirements of the floristics of the fertilizer type that will produce, plantation, product, the soil nutrient status etc. of using ground, this is that those skilled in the art can accomplish, does not preferably add chemical fertilizer during as the fertilizer of producing Organic food.
Combination azotobacter living bacteria count in described biological organic fertilizer or the biologic organic, inorganic composite fertilizer is preferably answered 〉=0.3 hundred million/gram when not containing phosphorus bacteria or potassium bacterium; When containing phosphorus bacteria and/or potassium bacterium, total living bacteria count is preferably answered 〉=0.6 hundred million/gram, and the living bacteria count of various function yeast is no less than 0.2 hundred million/gram; Water ratio generally is controlled in the 8-12% scope.
Characteristics of the present invention and beneficial effect:
1, bamboo plant rhizosphere association nitrogen fixation is by take the lead in an in the world research proposing and carry out of inventor herein.
2, after measured, has higher nitrogenase activity from the isolating described association nitrogen fixation bacterial strain of bamboo plant rhizosphere (WG-7 and WG-8); Owing to all separate, to the extremely well-adapted of bamboo rhizospheric environment from the bamboo plant rhizosphere.Studies have shown that for many years behind the inoculation bamboo plant, tool obviously improves bamboo root system and bamboo grove soil nitrogen fixing capacity, increases the bamboo leaves nitrogen content, promote the bamboo especially root growth of growing, improve effects such as bamboo shoot output and plant percent.Described bamboo forest biological fertilizer is after bamboo forest in use for bamboo shoot is used, and next year can be improved bamboo shoot output more than 16%.
3, because association nitrogen fixation has certain host specificity and homology, therefore its effect of bamboo forest biological fertilizer with the isolating combination azotobacter production of bamboo rhizosphere is that other types of biological fertilizer is irreplaceable in bamboo grove cultivation and operation.
4, be that the bamboo forest biological fertilizer produced of main bacteria seed is after having cooperated phosphate solubilizing bacteria, potassium solubilizing bacteria and other soil beneficial microorganism and fertilizer with bamboo rhizosphere combination azotobacter; not only can reduce or the use of alternative chemical fertilizer; but also has the soil micro-ecosystem of improvement environment; improve the soil water-reataining fertilizer-keeping performance; provide the bamboo plant growth required multiple efficacies such as various nutrients comprehensively, significant for production and the environment protection of green bamboo shoots and organic bamboo shoots.
5, bio-feritlizer in the market is various in style, and still, the special bio fertilizer of the suitable crop of matching species with the site is few.Owing to contain microorganism alive in the bio-feritlizer, and any biology all has limitation to natural adaptability, therefore it is impossible acting on all plants, all soil types, all weather condition with certain microbial fertilizer, and the microorganism that the suitable plant of matching species with the site is selected and the special bio fertilizer of production can have stronger specific aim to certain kind of plant, also have more the research application prospect.Improve the soil water-reataining fertilizer-keeping performance, provide the bamboo plant growth required multiple efficacies such as various nutrients comprehensively, significant for production and the environment protection of green bamboo shoots and organic bamboo shoots.
5, bio-feritlizer in the market is various in style, and still, the special bio fertilizer of the suitable crop of matching species with the site is few.Owing to contain microorganism alive in the bio-feritlizer, and any biology all has limitation to natural adaptability, therefore it is impossible acting on all plants, all soil types, all weather condition with certain microbial fertilizer, and the microorganism that the suitable plant of matching species with the site is selected and the special bio fertilizer of production can have stronger specific aim to certain kind of plant, also have more the research application prospect.
6, except associative nitrogen fixation, bacterial classification used in the present invention also has some indefinite growth-promoting functions to plant, as secreting growth stimulant class material and disease-resistant class material etc.
Embodiment
Embodiment 1
1) preparation vinelandii test tube solid slant culture base, its prescription is: glucose 0.5%, sucrose 0.5%, KH
2PO
40.04%, K
2HPO
4.3H
2O 0.01%, NH
4(SO
4)
20.1%, MgSO
4.7H
2O 0.02%, and NaCl 0.01%, CaCl
2.2H
2O 0.002%, FeCl
3.3H
2O 0.001%, Na
2MoO
4.2H
2O 0.0005%, K
2SO
40.15%, yeast extract 0.06%, agar 2%, pH6.8-7.2.After the sterilization, insert WG-7 and WG-8 bacterial strain respectively, 30～32 ℃ of cultivations make its surface growth full.
2) the shaking table culture medium of preparation vinelandii, its prescription is: glucose 0.5%, sucrose 0.5%, Semen Maydis powder 1%, K
2SO
40.15%, KH
2PO
40.04%, K
2HPO
4.3H
2O 0.01%, NH
4(SO
4)
20.1%, MgSO
4.7H
2O 0.02%, NaCl0.01%, CaCl
2.2H
2O 0.002%, FeCl
3.3H
2O 0.001%, Na
2MoO
4.2H
2O is mixed two kinds of liquid bio fertilizer by required volume ratio, just become hybrid bacterial strain liquid bio fertilizer.
4-2) the shaking table liquid culture with WG-7 and WG-8 bacterial strain inserts same fermentation cylinder for fermentation by required volume ratio, inoculum size 10%, leavening temperature 30-35 ℃, pH6.8-7.2,200 rev/mins of stirring velocitys, air flow 0.5～1: 1 cubic metre/1000 liters, incubation time 20～30 hours, make bacterial count reach 2,000,000,000/milliliter, also can be made into hybrid bacterial strain liquid bio fertilizer.
5) by aforesaid phosphorus bacteria substratum and present embodiment 1,2,3, the same quadrat method cultivation of 4-1 and the phosphorus bacteria that ferments, make phosphorus bacteria liquid bio fertilizer.
6) by aforesaid potassium bacteria culture medium and present embodiment 1,2,3, the same quadrat method cultivation of 4-1 and the potassium bacterium of fermenting, make potassium bacterial liquid bio-fertilizer.
7) fertile and liquid phosphorus bacteria bio-fertilizer and/or liquid potassium bacterium living beings fertilizer are mixed by required volume ratio with a certain combination azotobacter liquid bio of present embodiment 4-1 or 4-2 method preparation, just make the mixing liquid bio-fertilizer that contains combination azotobacter, phosphorus bacteria and/or potassium bacterium.
Embodiment 2
To be example, carry out the production of bamboo grove liquid bio fertilizer among the embodiment 1 from the isolating 2 strain combination azotobacters of bamboo plant rhizosphere.Also can carry out the production of bamboo grove liquid bio fertilizer with above-mentioned same method in the actual production only with any strain bacterium in the 2 strain bacterium.
Embodiment 3
The peat composed of rotten mosses is pulverized, crossed 80 mesh sieves, with 200 ℃ of hot blast cylinder drying and sterilizings, cool to room temperature.To adsorb with the peat composed of rotten mosses respectively with the various liquid bio fertilizer (comprising single culture and mixed strains) that embodiment 1 method is made, the viable count of its function yeast is reach the required standards, and make water content less than 30% (too high as water content, as can to adopt low temperature wind-force drying treatment) just make the various solid fungicides that contain single bacterial strain or hybrid bacterial strain.
Embodiment 4
With the animal excrement that ferment, as pig manure, through 200 ℃ of hot blast cylinder oven dry, sterilization; Again will be by the mixing liquid bio-fertilizer that contains combination azotobacter, phosphorus bacteria and/or potassium bacterium that the 7th step made among the embodiment 1, spray in the exsiccant animal excrement that fermented, bvg rolls and makes diameter 3～5mm particle on pan-pelletizer, cryodrying, make the living bacteria count 〉=0.6 hundred million/gram of combination azotobacter in every gram dried human and animal excreta material, phosphorus bacteria, potassium bacterium three sum, and biological organic fertilizer is made in the living bacteria count of every kind of bacterium 〉=0.2 hundred million/gram.
Embodiment 5
With the stalk that ferments and clay by weight 7: 3 ratio be mixed, pulverize, after the sterilization, adjust its nutrient content with fused(calcium magnesium)phosphate, Repone K, urea, make full N be about 3%, full P
2O
5Be about 3%, full K
2O is about 3%, the solid fungicide that to produce in embodiment 3 and ferment and the stalk of the clay that has been mixed is mixed with 4: 6 ratio and mixes thoroughly again, the living bacteria count of each function yeast is reach the required standards, add a certain amount of water, on pan-pelletizer, roll and make diameter 3～5mm particle, biologic organic, inorganic composite fertilizer is made in cryodrying.
Claims (3)
1, bamboo forest biological fertilizer, it is characterized in that being made of bamboo plant rhizosphere combination azotobacter and matrix, described bamboo plant rhizosphere combination azotobacter is from fixedly Paenibacillus polymyxa (Paenibacillus polymyxa) WG-7 bacterial strain and any bacterial strain in Bacillus licheniformis (Bacillus licheniformis) the WG-8 bacterial strain or the combination of two bacterial strains of molecular nitrogen the atmosphere of the isolating energy of bamboo plant rhizosphere.
2, bamboo forest biological fertilizer as claimed in claim 1, it is characterized in that this fertilizer also comprises phosphorus bacteria bacillus megaterium (Bacillus megaterium, culture presevation number is: ACCC10010) and/or potassium bacterium bacillusmusilaginosiengineering (Bacillus mucilaginosus, culture presevation number is: ACCC 10013).
3, bamboo forest biological fertilizer as claimed in claim 1 or 2 is characterized in that described matrix is a kind of of the existence of described bacterial classification or the liquid medium of cultivating, sorbent material, organic materials, inorganic material or several;
Described bio-fertilizer is: a) liquid bio fertilizer, b) solid fungicide, c) biological organic fertilizer and d) biologic organic, inorganic composite fertilizer any;
Described liquid bio fertilizer is the liquid bio fertilizer that the fermented liquid of described bacterial classification after the liquid nutrient medium fermentation forms;
Described solid fungicide is the solid fungicide that described liquid bio fertilizer forms after adsorbents adsorb; Or the freeze-drying microbial inoculum that after freeze-drying, forms; Or the dry microbial inoculum of spray-dried back formation.
Described biological organic fertilizer is the biological organic fertilizer of described liquid bio fertilizer or solid fungicide and described organic materials formation;
Described biologic organic, inorganic composite fertilizer is the biological organic and inorganic fertilizer that described liquid bio fertilizer or solid fungicide and described organic materials, inorganic material constitute;
Described sorbent material is a material: the mixture of one or more materials in the matter is planted in the peat composed of rotten mosses, peat, vermiculite, light calcium carbonate, kaolin, pond sludge, wilkinite, corruption;
Described organic materials is: a kind of material in the organic waste in crop material, wood chip, animal excrement, municipal wastes, mud, the industrial and agricultural production or the mixture of more than one materials; Or the organic fertilizer that behind fermentation maturity, forms again of above-mentioned organic materials.
Described inorganic material is a chemical fertilizer.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA021283710A CN1475467A (en) | 2002-08-14 | 2002-08-14 | Bamboo forest biological fertilizer |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA021283710A CN1475467A (en) | 2002-08-14 | 2002-08-14 | Bamboo forest biological fertilizer |
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| CN1475467A true CN1475467A (en) | 2004-02-18 |
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| CNA021283710A Pending CN1475467A (en) | 2002-08-14 | 2002-08-14 | Bamboo forest biological fertilizer |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101863720A (en) * | 2010-06-22 | 2010-10-20 | 河北冠祥生物科技开发有限公司 | Microbial fertilizer for holding water and fixing nitrogen of soil and preparation method thereof |
| CN105175195A (en) * | 2015-10-16 | 2015-12-23 | 北京盛大龙腾科技发展有限公司 | Composite growth promoter for promoting tobacco growth |
| CN105230297A (en) * | 2015-10-23 | 2016-01-13 | 上海金地农业发展股份有限公司 | Planting method of phyllostachys pracecox shoots |
| CN106748424A (en) * | 2016-11-30 | 2017-05-31 | 浙江科技学院 | Bamboo grove special bio charcoal base manure material, preparation method and applications |
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2002
- 2002-08-14 CN CNA021283710A patent/CN1475467A/en active Pending
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101863720A (en) * | 2010-06-22 | 2010-10-20 | 河北冠祥生物科技开发有限公司 | Microbial fertilizer for holding water and fixing nitrogen of soil and preparation method thereof |
| CN101863720B (en) * | 2010-06-22 | 2012-10-24 | 河北冠祥生物科技开发有限公司 | Microbial fertilizer for holding water and fixing nitrogen of soil and preparation method thereof |
| CN105175195A (en) * | 2015-10-16 | 2015-12-23 | 北京盛大龙腾科技发展有限公司 | Composite growth promoter for promoting tobacco growth |
| CN105175195B (en) * | 2015-10-16 | 2018-08-21 | 北京盛大龙腾科技发展有限公司 | A kind of composite growth-promoting agent promoting tobacco growing |
| CN105230297A (en) * | 2015-10-23 | 2016-01-13 | 上海金地农业发展股份有限公司 | Planting method of phyllostachys pracecox shoots |
| CN106748424A (en) * | 2016-11-30 | 2017-05-31 | 浙江科技学院 | Bamboo grove special bio charcoal base manure material, preparation method and applications |
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