CN101565689B - Production method for high-density pure arbuscular mycorrhizal fungal spore - Google Patents
Production method for high-density pure arbuscular mycorrhizal fungal spore Download PDFInfo
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- CN101565689B CN101565689B CN2009100989817A CN200910098981A CN101565689B CN 101565689 B CN101565689 B CN 101565689B CN 2009100989817 A CN2009100989817 A CN 2009100989817A CN 200910098981 A CN200910098981 A CN 200910098981A CN 101565689 B CN101565689 B CN 101565689B
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Abstract
The invention discloses a production method for high-density pure arbuscular mycorrhizal fungal spore, which belongs to the technical field of microorganism culturing, and comprises the following technical steps of: 1) culturing carrot root tissues converted from agrobacterium rhizogenes plasmid DNA; 2) culturing the carrot root tissues converted from agrobacterium rhizogenes plasmid DNA and glomus intraradices together in an improved synthetic culture medium; 3) taking a culture containing the root, hypha and spore infected by the arbuscular mycorrhizal fungus and transplanting the culture toa special culture box for culturing; 4) collecting the spore and mycelium; and 5) storing the arbuscular mycorrhizal fungal spore. Compared with the prior art, the production method for high-densitypure arbuscular mycorrhizal fungal spore has the advantages that: 1, the culture medium is an asepsis environment with low cost, which not only can meet the growing of a host plant, but also is suitable for the growing of AM epiphyte; 2, the nutrition requirements for the growing of the host plant are low and the host plant can grow and be cultured on the synthetic culture medium; 3, the spore cultured by the method is a non-pollution pure culture; and 4, the produced spore is stored in liquids and the activity can be maintained by above 90 percent.
Description
Technical field
The invention belongs to the microorganism culturing technical field, be specifically related to a kind of purebred working method of high-density of bush mycorrhizal fungi spore.
Background technology
(Arbuscular Mycorrhizae is that occurring in nature is ubiquitous AM) to arbuscular mycorrhiza, with the symbiotic fungi of 80% above land plant.At occurring in nature, host of AM fungi and distribution range are all very extensive, can form symbiotic relationship with most plants, have found that so far more than 150 kind of AM fungi can form symbiote with plant, are one of types the most general in the mycorhiza.It and host plant reciprocal symbiosis: plain and other nutritive element of the outer mycelia of the root of fungi Absorption of Phosphorus from soil is supplied with host plant; Improved the receptivity of plant to mineral element; Promote plant-growth etc., host plant is then supplied with the necessary glucide of fungal growth.Because the critical role of AM fungi in plant life, simultaneously as a kind of biotechnology measure, its application prospect at aspects such as eagroforestry, greening and environment protection is very wide, and very high expectation has all been expressed to its application in countries in the world.Therefore, arbuscular mycorrhiza progressively develops into for a ten minutes active research field, and Chinese scholars has carried out exploring widely and research to aspects such as its resource, classification, physiological ecological and application thereof.
At present developed country such as America, Europe, Asia, the Pacific Ocean and developing country are all stepping up to study the AM mycorrhizal fungi at Application for Field approaches and methods such as eagroforestry, greening and environment protection, have obtained progress faster in recent years.Because the AM fungi is the obligate symbiosis fungi, self can not synthetic DNA, up to the present, the pure culture of this fungi is not achieved success as yet, so can not as the ectomycorrhiza fungi, produce the inoculation microbial inoculum on a large scale through industrial fermentation.Can only it be seeded on the root system of plant, the symbiosis culture through 2-4 month grows with host plant, just can obtain its propagulum.The method that the AM microbial inoculum is produced, summarize and get up to have following several kinds: potted plant culture method, static culture method, flow culture method, spraying culture method, land for growing field crops culture method etc., wherein potted plant culture method is in the method that is still a kind of traditional economy at present.The production application commercialization of at present more existing national fungi preparations; All there is the AM microbial inoculum to sell like states such as the U.S., Britain, France, Japan; Their preparation purity is high, and quality is good, mostly controls various environmental factors through strictness and carries out greenhouse batch production production; Produce AM microbial inoculum by name like farming research center, Britain Lausanne and apply to vegetables, flowers, fruit tree etc. widely in Britain, France, Denmark, Holland and Japan and other countries; In nz and the production of Australian herbage, remarkable economic efficiency and ecological benefits have all been obtained simultaneously.But China does not also have the commercial microbial inoculum production of batch production at present, so the high-efficient culture technology of AM fungi mycorhiza all has profound significance to the eagroforestry of China, environment protection etc.
The conventional cultural method of bush mycorrhizal fungi mainly is that employing soil is pushed up or river sand is matrix, and monocrop (like corn, trifolium) is a host plant.The culture efficiency of mycorrhizal fungi can be improved through composition and the culture condition of regulating culture medium; Atimanav etc. have compared influence (the At imanav Gaur of the expansion mineral material of different-grain diameter to bush mycorrhizal fungi production; A.dholeya.Effects of the particlesize of soil-less substrates upon AM fungus inoculum production 2000,10:43-48); Britain PLANT WORK company (www.Plantworksuk.uk) adopts a kind of expansion mineral to cultivate bush mycorrhizal fungi as matrix, in culturing process, has used the growth of the control of slow-releasing phosphate fertilizer and adjusting plant and mycorrhizal fungi simultaneously.These methods adopt single expansion mineral or expansion mineral to add the mode of careless carbon mostly, and culture efficiency is not high enough.Document (Sylvia; D.M.and D.H.Hubbell.1986.Growthand sporulation of vesicular-arbuscular mycorrhizal fungi in aeroponicand membrane systems.Symbiosis 1:259-267) adopt aeroponics to cultivate microbial inoculum; In airtight container, nutritive medium is become aerosol and be sprayed on and infect on the root system of plant that mycorrhizal fungi is arranged, obtain purified bacterium chaste tree with the atomizing assembling device.The advantage of this method is to obtain very purified microbial inoculum, but cost high, yield poorly.
It all is to be employed under the greenhouse experiment host plant and individual month mode of bush mycorrhizal fungi co-cultivation 4-5 are carried out that the bush mycorrhizal fungi microbial inoculum is commercially produced; Because bush mycorrhizal fungi can not leave the host plant single culture; Therefore this microbial inoculum scale operation ten minutes difficulty, and the type of host plant, the prescription of culture medium, the key that culture condition is production technique.
Summary of the invention
To the problem that prior art exists, the object of the invention be to provide a kind of can scale operation, cost is low, the technical scheme of the purebred working method of high-density of the bush mycorrhizal fungi spore that output is high.
The purebred working method of the high-density of described bush mycorrhizal fungi spore is characterized in that comprising following process step:
The root tissue of the Radix Dauci Sativae root that 1) the Agrobacterium rhizogenes DNA is transformed is cultivated in improving synthetic medium, in the dark, under the temperature 23-25 ℃ of condition, cultivates 5-7 week;
The root tissue of the Radix Dauci Sativae root that 2) the Agrobacterium rhizogenes DNA of above-mentioned cultivation is transformed is forwarded in the new improvement synthetic medium; Again at the inoculation of medium bush mycorrhizal fungi; Bush mycorrhizal fungi inoculating spores amount is 300-500/mL; In the dark, under the temperature 23-25 ℃ of condition, cultivate 7-10 week;
3) in step 2) substratum in to get the diameter that contains the root, mycelia and the spore that are infected by bush mycorrhizal fungi be that culture culture transferring to special-purpose the cultivation in the box of 1.5-2.5cm contained a Room of improving synthetic medium; And the root tissue culture transferring of the Radix Dauci Sativae root that transforms the Agrobacterium rhizogenes DNA of fresh culture is by culture; The described special-purpose box of cultivating is that box is cultivated in two Room; 350-450 purpose tabula net is set between two Room, and special-purpose the cultivation in another chamber of box contained bush mycorrhizal fungi RT substratum, with above-mentioned culture in the dark; Under the temperature 23-25 ℃ of condition, cultivate 7-10 week;
4) get the above-mentioned special-purpose box of cultivating and contain spore and mycelium in bush mycorrhizal fungi RT substratum one Room; Be dissolved in the aseptic sodium citrate soln of 8-11 mmole; And accent pH to 5.5-6.5; Fully stir, be poured into the sieve in aseptic 0.45um aperture again, wash extremely only surplus spore and mycelium with sterilized water;
5) get above-mentioned spore that obtains and mycelium, collect spore, after cleaning up, be soaked in 20% the glycerine sterile solution that contains Streptomycin sulphate 100-300mg/L and qingfengmeisu qiong 50-200mg/L and preserve with sieve.
The mycorrhizal fungal spores purebred production methods, high-density, wherein said improvement synthetic medium containing: magnesium 720-740mg / L, potassium nitrate 70-90mg / L, potassium chloride 55 - 75mg / L, calcium nitrate 270-290mg / L, EDTA iron sodium 5-12mg / L, inositol 40-60mg / L, manganese sulfate 0.01-0.08mg / L, copper sulfate 0.01-0.03mg / L, boric acid 0.3 -0.6mg / L, zinc sulfate 0.1-0.4mg / L, potassium iodide, 0.01-0.03mg / L, sodium molybdate 0.01-0.03mg / L, cobalt chloride, 0.01-0.04mg / L, vitamin B0.1-0.3mg / L, pyridoxine BEEP 0.1-0.3mg / L, nicotinic acid 0.2-0.8mg / L, sucrose 7500-8500mg / L, glucose 4500-5500mg / L and vegetable gum 3500-4500mg / L.
The mycorrhizal fungal spores purebred production methods, high-density, wherein said improvement synthetic medium containing: magnesium 725-735mg / L, potassium nitrate 75-85mg / L, potassium chloride 60 - 70mg / L, calcium nitrate 275-285mg / L, EDTA iron sodium 7-9mg / L, inositol 45-55mg / L, manganese sulfate 0.01-0.05mg / L, copper sulfate 0.01-0.02mg / L, boric acid 0.4 -0.5mg / L, zinc sulfate 0.2-0.3mg / L, potassium iodide, 0.01-0.02mg / L, sodium molybdate 0.01-0.02mg / L, cobalt chloride, 0.02-0.03mg / L, vitamin B0.1-0.2mg / L, pyridoxine BEEP 0.1-0.2mg / L, nicotinic acid 0.4-0.6mg / L, sucrose 7800-8200mg / L, glucose 4800-5200mg / L and vegetable gum 3700-4200mg / L.
The purebred working method of the high-density of described bush mycorrhizal fungi spore; It is characterized in that step 2) in bush mycorrhizal fungi be the Paraglomus of bush mycorrhizal fungi; The amount of bush mycorrhizal fungi inoculating spores is 350-450/mL; Culture temperature is under the 23.5-24.5 ℃ of condition, cultivates 8-9 week.
The purebred working method of the high-density of described bush mycorrhizal fungi spore is characterized in that step 2) in bush mycorrhizal fungi be that sacculus is mould in the root.
The purebred working method of the high-density of described bush mycorrhizal fungi spore is characterized in that the special-purpose tabula net of cultivating the middle setting of box is the 370-400 order in the step 3), and culture condition is: in the dark, under the temperature 23.5-24.5 ℃ of condition, cultivate 8-9 week.
The purebred working method of the high-density of described bush mycorrhizal fungi spore; It is characterized in that bush mycorrhizal fungi RT substratum is minimizing sucrose and two components of glucose in improving synthetic medium in the step 3), and add the tartrate ammonia of substratum weight 0.1-0.2% and the root leach liquor of culture volume 2-5%.
The purebred working method of the high-density of described bush mycorrhizal fungi spore; It is characterized in that the special-purpose box of cultivating of step 4) contains spore and mycelium in bush mycorrhizal fungi substratum one Room; Be dissolved in the aseptic sodium citrate soln of 9-10 mmole, and transfer pH to 5.8-6.2.
The purebred working method of the high-density of described bush mycorrhizal fungi spore is characterized in that in the step 5) containing Streptomycin sulphate 150-250mg/L and qingfengmeisu qiong 100-150mg/L in 20% the glycerine sterile solution.
The purebred working method of the high-density of described bush mycorrhizal fungi spore is characterized in that the liquid that the root leach liquor obtains for the Radix Dauci Sativae root tissue of cultivating the conversion of Agrobacterium rhizogenes DNA.
The present invention improves synthetic medium and adopts special-purpose cultivation box to carry out the cultivation of bush mycorrhizal fungi through configuration; Compared with prior art; The invention has the advantages that: 1 substratum is a gnotobasis, and is with low cost, not only satisfies the host plant growth but also be fit to the AM fungal growth; It is low that 2 host plant growing nutrients require, can be on synthetic medium grown cultures; 3 spores (containing mycelium) by this method cultivation are pollution-free axenic cultures; The spore of 4 productions is stored in the liquid, can keep more than 90% activity.The spore that present technique is produced can be used for the breeding of bush mycorrhiza agent, the production of bio-feritlizer, and free of contamination purebred spore is provided in the scientific research, be used for fundamental research.
Embodiment
Below further specify the present invention through specific embodiment.Bush mycorrhizal fungi is all on sale on market, and the technology of the Radix Dauci Sativae root that the Agrobacterium rhizogenes DNA transforms is prior art.
Embodiment:
1. substratum preparation
Be used to cultivate the Radix Dauci Sativae root of Agrobacterium rhizogenes DNA conversion and the substratum composition of bush mycorrhizal fungi (concentration of component is mg/L, with the preparation of ultra-clean water):
Sal epsom 731 saltpetre 80
Repone K 65 nitrocalcite 288
EDTA ferrisodium salt 8 inositols 50
Manganous sulfate 0.04 copper sulfate 0.025
Boric acid 0.45 zinc sulfate 0.22
Potassiumiodide 0.01 Sodium orthomolybdate 0.02
NSC 51149 0.025 vitamins B 0.1
BEEP pyridoxine? 0.1? Niacin 0.5
Sucrose 8000 glucose 5000
Wherein Mn and Zn are controlled at low-down concentration.These two kinds of elements can suppress the sprouting of spore.Substratum adds 4000mg/L vegetable jelly (account for substratum weight 0.4%) and solidifies, because contain high phosphorus concentration in the vegetable jelly, so in substratum, save KH
2PO
4Substratum is at 1 kilogram of pressure, 121 ℃ of sterilization 20min.The preparation of bush mycorrhizal fungi RT substratum is exactly in improving synthetic medium, to reduce sucrose and these two components of glucose and add tartrate ammonia and the root leach liquor.
2. root tissue is cultivated
The Radix Dauci Sativae root that the Agrobacterium rhizogenes DNA is transformed is cultivated in as above substratum, in the dark, and under 24 ℃ of the temperature; Cultivated for 5 week, be cultured to root tissue and cover with whole petridish (15cm diameter), again root tissue is forwarded in the new improvement synthetic medium; Sacculus is mould in the inoculation of medium root simultaneously, and inoculum size is 400/ml, in the dark; Under 24 ℃ of the temperature, cultivated for 8 week.
3. culture transferring and sporogenesis
The culture of getting a fritter that contains the root, mycelia and the spore that are infected by bush mycorrhizal fungi in the above-mentioned substratum moves and is connected to the wherein Room that two Room stainless steels are cultivated box; Contain the improvement synthetic medium in this chamber, simultaneously the bright new root tissue culture transferring of cultivating to this fritter inoculum next door; And bush mycorrhizal fungi RT substratum is contained in an other Room of petridish; Bush mycorrhizal fungi RT substratum contains the tartrate ammonia of substratum weight 0.1% or 0.2% and the root leach liquor (containing the sesquiterpene element) of culture volume 2%, 3% or 5%, and middle tabula net 400 orders do not allow root growth to this chamber.So in the dark, after 24 ℃ of temperature cultivated for 8 week down, tabula was in another chamber procreation growth in the middle of hypha,hyphae just got over.
4. spore is gathered in the crops
Spore after containing another chamber culture medium culturing maturation of bush mycorrhizal fungi RT substratum is dissolved in together with substratum in the aseptic sodium citrate soln of 10mM; And accent pH to 6.0; In magnetic stirring apparatus, fully stir and make the solid gums dissolving; Be poured into again on the sieve in aseptic 0.45um aperture, wash to having only spore and mycelium with sterilized water.
5. spore is preserved
Collect spore with sieve, after cleaning up, just can be soaked in the sterile solution that contains 20% glycerine, Streptomycin sulphate (200mg/L), qingfengmeisu qiong (100mg/L), can preserve more than 1 year under 4 ℃.
Spore quantity detects and can adopt stereoscopic microscope to amplify 20 times of observation countings.It is 1000/ml that product can be divided into spore quantity, three different sizes of 5000/ml and 10000/ml, and the spore count 100-120 that obtains of the technology before the present invention/ml, and the spore count that technology of the present invention obtains is generally at 1500-2000/ml.
The content of substratum is (unit is mg/L) in the step 1:
720 magnesium sulfate, potassium nitrate 70 55 Potassium nitrate, calcium 270, EDTA sodium iron 5, inositol 40 0.01 manganese sulfate, copper sulfate 0.01 0.3 boric acid, zinc sulfate, 0.1, potassium iodide 0.01 0.01 sodium molybdate, cobalt chloride 0.01, vitamin B0.1, 0.1 BEEP pyridoxine, niacin 0.2, 7500 sucrose, glucose, vegetable gum 4500 and 3500
Or 740 using magnesium sulfate, potassium nitrate, 90, 75 potassium chloride, calcium nitrate, 290, EDTA sodium iron 12, inositol 60 0.08 manganese sulfate, copper sulfate 0.03 0.6 boric acid, zinc sulfate, 0.4, 0.03 potassium iodide, sodium molybdate 0.03, 0.04 cobalt chloride, vitamin B0.3, 0.3 BEEP pyridoxine, niacin 0.8, sucrose and glucose 8500 5500;
Or 725 using magnesium sulfate, potassium nitrate 85 60 Potassium nitrate, calcium 275, EDTA sodium iron 7, inositol 45 0.01 manganese sulfate, copper sulfate 0.01 Boric acid 0.4 0.2 zinc sulfate, potassium iodide 0.015 Sodium molybdate 0.02, 0.02 cobalt chloride, vitamin B0.2, BEEP pyridoxine 0.2, niacin 0.6, sucrose and glucose 5200 8200 4500 vegetable gum.
The Paraglomus that the bush mycorrhizal fungi of step 2 changes bush mycorrhizal fungi into gets other fungies; Culture condition adopts in the dark; 23,23.5,24,24.5 or 25 ℃ of temperature were cultivated for 7,8,9 or 10 weeks down, and the spore inoculating amount is 300/ml, 350/ml, 400/ml or 450/ml, and culture condition adopts in the dark in the step 3; Temperature was cultivated 50,55 or 60 days down for 23,23.5,24,24.5 or 25 ℃, and the special-purpose box tabula net of cultivating adopts 350 or 370 orders; Adopt in the step 4 in the aseptic sodium citrate soln of 8mM, 9mM or 11mM, and transfer pH to 5.5,6.0 or 6.5; Add Streptomycin sulphate 100mg/L, qingfengmeisu qiong 50mg/L in the step 5; Or Streptomycin sulphate 300mg/L, qingfengmeisu qiong 200mg/L; Or Streptomycin sulphate 150mg/L, qingfengmeisu qiong 150mg/L, other steps are all identical with embodiment with condition, also can reach the technique effect identical with embodiment at last.
Claims (7)
1. the purebred working method of the high-density of bush mycorrhizal fungi spore is characterized in that comprising following process step:
The root tissue of the Radix Dauci Sativae root that 1) the Agrobacterium rhizogenes DNA is transformed is cultivated in improving synthetic medium, in the dark, under the temperature 23-25 ℃ of condition, cultivates 5-7 week;
The root tissue of the Radix Dauci Sativae root that 2) the Agrobacterium rhizogenes DNA of above-mentioned cultivation is transformed is forwarded in the new improvement synthetic medium; Again at the inoculation of medium bush mycorrhizal fungi; Bush mycorrhizal fungi inoculating spores amount is 300-500/mL; In the dark, under the temperature 23-25 ℃ of condition, cultivate 7-10 week;
3) in step 2) substratum in to get the diameter that contains the root, mycelia and the spore that are infected by bush mycorrhizal fungi be that culture culture transferring to special-purpose the cultivation in the box of 1.5-2.5cm contained a Room of improving synthetic medium; And the root tissue culture transferring of the Radix Dauci Sativae root that transforms the Agrobacterium rhizogenes DNA of fresh culture is by culture; The described special-purpose box of cultivating is that box is cultivated in two Room; 350-450 purpose tabula net is set between two Room, and special-purpose the cultivation in another chamber of box contained bush mycorrhizal fungi RT substratum, with above-mentioned culture in the dark; Under the temperature 23-25 ℃ of condition, cultivate 7-10 week;
4) get the above-mentioned special-purpose box of cultivating and contain spore and mycelium in bush mycorrhizal fungi RT substratum one Room; Be dissolved in the aseptic sodium citrate soln of 8-11 mmole; And accent pH to 5.5-6.5; Fully stir, be poured into the sieve in aseptic 0.45um aperture again, wash extremely only surplus spore and mycelium with sterilized water;
5) get above-mentioned spore that obtains and mycelium, collect spore, after cleaning up, be soaked in 20% the glycerine sterile solution that contains Streptomycin sulphate 100-300mg/L and qingfengmeisu qiong 50-200mg/L and preserve with sieve;
Synthetic medium containing the above-mentioned improvements: Magnesium sulfate 720-740mg / L, potassium 70-90mg / L, potassium chloride 55-75mg / L, calcium nitrate 270-290mg / L, EDTA iron sodium 5-12mg / L , inositol 40-60mg / L, manganese sulfate 0.01-0.08mg / L, copper sulfate 0.01-0.03mg / L, boric acid 0.3-0.6mg / L, zinc sulfate 0.1-0.4mg / L, potassium iodide, 0.01-0.03mg / L, sodium molybdate 0.01-0.03mg / L, cobalt chloride, 0.01-0.04mg / L, vitamin B0.1-0.3mg / L, pyridoxine BEEP 0.1-0.3mg / L, nicotinic acid 0.2-0.8mg / L , sucrose 7500-8500mg / L, glucose 4500-5500mg / L and vegetable gum 3500-4500mg / L;
Above-mentioned bush mycorrhizal fungi RT substratum is in improving synthetic medium, to reduce sucrose and two components of glucose; And the tartrate ammonia of adding substratum weight 0.1-0.2% and the root leach liquor of culture volume 2-5%, the described liquid that leach liquor obtains for the Radix Dauci Sativae root tissue of cultivating the conversion of Agrobacterium rhizogenes DNA.
(2) as claimed in claim 1, wherein the arbuscular mycorrhizal fungal spores producing pure high density, wherein the improvement comprising synthetic medium: Magnesium sulfate 725-735mg / L, potassium 75-85mg / L , potassium chloride, 60-70mg / L, calcium nitrate 275-285mg / L, EDTA iron sodium 7-9mg / L, inositol 45-55mg / L, manganese sulfate 0.01-0.05mg / L, copper sulfate 0.01-0.02 mg / L, boric acid 0.4-0.5mg / L, zinc sulfate 0.2-0.3mg / L, potassium iodide, 0.01-0.02mg / L, sodium molybdate 0.01-0.02mg / L, cobalt chloride, 0.02-0.03mg / L, vitamins B0.1-0.2mg / L, pyridoxine BEEP 0.1-0.2mg / L, nicotinic acid 0.4-0.6mg / L, sucrose 7800-8200mg / L, glucose 4800-5200mg / L and vegetable gum 3700-4200mg / L.
3. the purebred working method of the high-density of bush mycorrhizal fungi spore as claimed in claim 1; It is characterized in that step 2) in bush mycorrhizal fungi be the Paraglomus of bush mycorrhizal fungi; The amount of bush mycorrhizal fungi inoculating spores is 350-450/mL; Culture temperature is under the 23.5-24.5 ℃ of condition, cultivates 8-9 week.
4. the purebred working method of the high-density of bush mycorrhizal fungi spore as claimed in claim 1 is characterized in that step 2) in bush mycorrhizal fungi be that sacculus is mould in the root.
5. the purebred working method of the high-density of bush mycorrhizal fungi spore as claimed in claim 1; It is characterized in that the special-purpose tabula net of cultivating the middle setting of box is the 370-400 order in the step 3); Culture condition is: in the dark, under the temperature 23.5-24.5 ℃ of condition, cultivate 8-9 week.
6. the purebred working method of the high-density of bush mycorrhizal fungi spore as claimed in claim 1; It is characterized in that the special-purpose box of cultivating of step 4) contains spore and mycelium in bush mycorrhizal fungi substratum one Room; Be dissolved in the aseptic sodium citrate soln of 9-10 mmole, and transfer pH to 5.8-6.2.
7. the purebred working method of the high-density of bush mycorrhizal fungi spore as claimed in claim 1 is characterized in that in the step 5) containing Streptomycin sulphate 150-250mg/L and qingfengmeisu qiong 100-150mg/L in 20% the glycerine sterile solution.
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| CN102652957A (en) * | 2012-05-08 | 2012-09-05 | 江苏大学 | Remediation method of soil polycyclic aromatic hydrocarbon by combining surfactant producing bacteria and mycorrhiza |
| CN103981100A (en) * | 2014-05-15 | 2014-08-13 | 浙江师范大学 | Method for promoting sprouting of arbuscular mycorrhizal fungi spore and growth of hypha |
| CN104025983A (en) * | 2014-06-06 | 2014-09-10 | 浙江师范大学 | Application of dual inoculation of arbuscular mycorrhiza fungi and rhizobium for promoting leguminous plants to absorb phosphorus minerals |
| CN109207370B (en) * | 2017-07-06 | 2021-08-24 | 上海臻衍生物科技有限公司 | Complex culture medium and symbiotic inclusion compound of arbuscular mycorrhizal fungi and application thereof |
| CN109197385B (en) * | 2018-09-20 | 2021-07-02 | 浙江世佳科技股份有限公司 | Arbuscular mycorrhizal fungi culture medium |
| CN110150324A (en) * | 2019-07-03 | 2019-08-23 | 安徽新熙盟生物科技有限公司 | A kind of compound nitragin, preparation method and application |
| CN111909855A (en) * | 2020-07-23 | 2020-11-10 | 华南农业大学 | Application of gibberellin-deficient mutant in promoting arbuscular mycorrhizal fungi spore production |
| CN114774289A (en) * | 2022-04-13 | 2022-07-22 | 安徽农业大学 | Method for improving spore production quality and efficiency of arbuscular mycorrhizal fungi |
| CN115029293A (en) * | 2022-06-08 | 2022-09-09 | 南京大学 | Method for efficiently propagating ascosphaera radiculosa spores through hairy roots of carrots |
| CN114933978B (en) * | 2022-07-25 | 2022-11-04 | 烟台泓源生物肥料有限公司 | Arbuscular mycorrhizal fungi culture medium |
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