CN106508429A - Multiplication method of arbuscular mycorrhiza fungi inoculant in coal mine area - Google Patents
Multiplication method of arbuscular mycorrhiza fungi inoculant in coal mine area Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
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Abstract
The invention discloses a multiplication method of arbuscular mycorrhiza (AM) fungi inoculant in a coal mine area and provides a preparation method of the AM fungi inoculant. The multiplication method comprises the following steps that A, a host plant of AM fungi is cultivated in a cultivation medium to which the AM fungi is applied, the root system of the cultivated host plant and the cultivation medium are collected, and thus the AM fungi inoculant is obtained. Experiments prove that by the adoption of a pot cultivation method, the defects that existing multiplication methods are long in cultivation period, high in medium cost and unstable in quality are overcome. The AM fungi inoculant multiplied by the method has the following advantages that media like decomposed coal and sandy soil are low in cost and easy to obtain, quality is light, transportation is easy, the period is short, the infection degree is high, and the quality of the inoculants is high. The formula of the AM fungi inoculants provides technical guidance for multiplication of high-quality AM fungi inoculants in the coal mine area.
Description
Technical field
The present invention relates to the production method of microbial bacterial agent, more particularly, to a kind of coal field bush mycorrhizal fungi preparation
Expanding propagation method.
Background technology
The symbiosis that mycorhiza (Mycorrhiza) is mycorrhizal fungi to be formed with root system of plant, more than 93% plant in nature
Mycorhiza can be formed.AMF is also known as AM fungies (Arbuscular mycorrhiza Fungi, abbreviation AMF).Forefathers
Research thinks that AM fungies can promote plant to mineral nutrient especially phosphorus and micro- absorption, strengthen plant resistant and support
Divide the ability of stress, reduce nitrogen, the usage amount of phosphate fertilizer and loss, so as to avoid body eutrophication.Meanwhile, its mycelia can be with shape
Into Hyphal links, nutrient is transmitted between different plants, become the important channel of ecosystem nutrient circulation.AM fungies can improve plant
The resistance of thing：The water regime of plant can be improved, the drought-resistant ability of plant is improved；The ability of plant anti-salt stress can be improved；
Genes For Plant Tolerance organic contamination ability can be improved；The resistance against diseases of plant can be improved；Resisting for plant heavy metal pollution can be improved
Ability.
In view of beneficial effect of the AM fungies in terms of agriculture and forestry and revegetation, exploitation AMF " bio-feritlizer "
Very promising, its primary problem for solving is exactly to realize the high-purity culture of AM fungies.Pure culture ideal at present
The cultural method of method AM fungal inoculant mainly have potted plant cultivation, a bead locellus method, bead locellus cultivation it is excellent
Point is can to separate culture matrix and AM fungies, obtains pure AM mycothalluses, but which is relatively costly, and production technology is not
Enough maturations, only for scientific research and the production and application of small-scale Inoculant；Live body host plant basin carries cultivation culture AM fungies
Advantage be reliability convenient and simple for operation, have the disadvantage that cultivation cycle length, quality is unstable, brood body yield is few, but at present it
It is still the main method of AM fungal inoculant cultures.With reference to arbuscular mycorrhiza (AM) fungi pure culture research method current research into
Really, the aseptic dual in vitro culture method of locellus Ri-T-DNA transition Carrot Roots of AM fungies monospore, this method is improved,
The production method of the AM fungal spore microbial inoculums of presently the most highly effective and safe can be obtained, but containers demand is high, technology content is higher, only
For scientific research, however, large-scale microbial inoculum production will be realized, need to expand numerous under the conditions of land for growing field crops (outdoor opening).
Relevant arbuscular mycorrhiza expands the culture that numerous disclosed patented technology content pertains only to AM fungi broad spectrum activities mostly at present
Method (for example, develop specific culture apparatus, using the culture matrix of hybridization) and application of the AM fungies in farmland production
(for example, a certain AM fungies are realized the efficient field planting of plant, plant improvement of roasting side's note etc.).
" ten thousand methods of production bush mycorrhizal fungi preparation " of Patent No. CN1511944A, relate generally to a kind of using mixing
Culture matrix, the method for selecting the mode that various host plant mixed seeding are mixed up to produce bush mycorrhizal fungi preparation, Patent No.
A kind of " efficient drought-resistant, high phoshpate-tolerant nutrition group planting mycorrhizal fungi and its production method " of CN1548524A, relates generally to a kind of choosing
With Chinese sorghum as host plant, using zeolite, river sand mixture as culture matrix producing efficient drought-resistant, high phoshpate-tolerant nutrition
The production method of AMF Glomus mosseae (Glomus mosseae) 93-6 bacterial strains；And Patent No.
A kind of " method for improving corn yield " of CN103329743, relates generally to by being inoculated with to corn under the conditions of separate furrow irrigation
AMF Glomus versiforme (G1omus verszforme) microbial inoculum, discloses a kind of method for improving corn yield.Specially
" culture from mycorrhizal fungi with bead as culture matrix " of the profit number for CN1354252A, relates generally to a kind of using special
Fixed culture apparatus and culture medium carry out the cultural method of AMF；" one kind of Patent No. CN104371932A.
The production method of AM fungal inoculants ", mainly by Glomus mosseae, mould contracting sacculus, Glomus intraradices and Glomus versiforme four
The expansion for planting AM fungies is numerous, produces high-quality AM fungal inoculants, and by realizing the mycorrhizal seedling raising that structure is offered sacriffices to the gods or the spirits of the dead from numerous microbial inoculum,
The land for growing field crops inoculation of potato and celery, achieves very good effect.
Usually need to buy culture matrix using said method, be difficult with localizing the matrix being easy to get and effectively expanded
It is numerous, it is not strong to ecologically fragile areas application, at present, still there is no selectivity, the multiplication technique of standardized mining area localization.Cut-off
2010, the damage area of China's coal-mine exploitation reached national strip coal mining and has destroyed 30,000 hm of the soil gross area2More than；China
Strip mining transformation is used mostly outer dump mode, and it is destroy soil amount 1.5 2.0 times which covers area, and the whole nation is accumulative to be taken
About 5.6 ten thousand hm of soil2, hence it is imperative that setting up its special AM fungies multiplication technique and method for mining area, it is mining area soil
Reclaim to provide with ecological recovery and technological guidance and establish application foundation.
The content of the invention
The good method of drawbacks described above, the present invention is overcome to carry out basin using different substrates proportioning and carry cultivation, overcome at present
Existing method cycle length, matrix are difficult to obtain, the defect of high cost, there is provided a kind of expansion of coal field bush mycorrhizal fungi preparation
Breeding method, the environmentally friendly matrix that is easy to get, microbial inoculum small volume, quality transport storage, microbial inoculum purity height easily, the AM fungies of the present invention
Microbial inoculum is filled a prescription, and is especially the numerous offer technological guidance of expansion of coal field high-quality AM microbial inoculum.
It is an object of the present invention to provide a kind of preparation method of AM fungal inoculants.
The method that the present invention is provided, comprises the steps A：By the host plant of AM fungies in the culture for being applied with AM fungies
Cultivate in matrix, collect the root system and culture matrix of the host plant after cultivating, that is, obtain AM fungal inoculants；
The culture matrix for it is following 1) or 2)：
1) culture matrix shown in is made up of sandy soil, vermiculite and perlite；
2) culture matrix shown in is made up of sandy soil, vermiculite, perlite and weathered coal.
In said method, 1) shown in culture matrix in, the sandy soil, the vermiculite and the perlitic volume ratio are
2-3:1:1；
2), in the culture matrix shown in, the volume ratio of the sandy soil, the vermiculite, the perlite and the weathered coal is
1-2:1:1:1。
In said method,
After the collection culture, the root system of host plant and the time of culture matrix are to form host after per gram of culture
When the AM fungal spore quantity of the root system and culture matrix of plant reaches the amount of 40-50；
Or the incubation time is no less than 3 months；
Or the incubation time is 3-5 month.Or the incubation time is 3 months.
In said method, in methods described, also include the step of host plant seed is carried out into vernalization before culture.
In said method, the condition of the vernalization is 28-30 DEG C of light culture 3 days.
In said method, the host plant is can be with the plant of AM mycosymbiosises；
Or, described can be corn, Chinese sorghum, clover with the plant of AM mycosymbiosises.
In said method, the AM fungies are Moses's pipe handle capsule mould or Glomus intraradices bacterium.
In said method, the corresponding culture matrix of Moses's pipe handle capsule mould for it is described 2) shown in culture matrix；
The corresponding culture matrix of the Glomus intraradices bacterium for it is described 1) shown in culture matrix.
Another object of the present invention is to provide a kind of matrix suitable for coal field cheap and easy to get, small volume, light weight, easily
The high AM fungal inoculant expanding propagation methods of fortune storage, microbial inoculum purity.
The method that the present invention is provided includes step A in said method, realizes that AM fungal inoculants expand numerous.
The experiment proves that, the present invention carries cultivation using basin, overcomes existing cultivation cycle length, matrix and is difficult
The high defect of procurement cost, the inventive method expand numerous AM fungal inoculants and have the advantage that：
1) environmentally friendly matrix：The perlite of present invention employing, vermiculite wind cheap and easy to get, high with coal field organic matter
Change coal to combine, small volume, easy shipping storage；Weathered coal, sandy soil etc. belong to coal field common materials, turn waste into wealth, and improve
Resource utilization, meets the fundamental state policy for economizing on resources, for vast propagation, is particularly suited for coal field ecology and repaiies
It is multiple, it is also widely applied to agriculture and forestry development；
2) cycle is short, material are common：The production method of the AM fungal inoculants that the present invention is provided, which includes matrix sterilizing, jade
Meter Cui Ya, inoculation, seedling management, microbial inoculum quality analysis；By (Glomus mo sseae, abbreviation Gm) mould to Moses's pipe handle capsule,
The expansion of Glomus intraradices (Glomus intraradices, write a Chinese character in simplified form Gi) bacterial classification is numerous, and the AM fungal inoculants produced infect degree
Height, spore amount are big.
3) infection strength is high
Glomus intraradices to the mycorhiza infection strength of corn more than 72%, invade by the mould mycorhiza to corn of Moses's pipe handle capsule
, more than 91%, Gm, Gi are in sandy soil 2 for dye intensity：Vermiculite 1：Mycorhiza in the composition matrix of perlite 1 (Z formulas) to corn
What infection strength studied pure sandy soil (S formulas) than ever has been respectively increased 6.64%, 15.51%, and Gm, Gi are in sandy soil 1: vermiculite 1:
Perlite 1：Study dividing for pure sandy soil (S formulas) in the matrix of weathered coal 1 (W formulas) to the mycorhiza infection strength of corn than ever
6.72%, 8.19% is not improve.
4) microbial inoculum quality is high
Compared with the spore density of pure sandy soil, matrix W formula Moses's pipe handle capsule is mould, Glomus intraradices are invaded to the mycorhiza of corn
Dye intensity study than ever pure sandy soil S formulation medias be respectively increased 32/g, 4/g, matrix Z Moses's pipe handle capsule is mould, root
The mould mycorhiza infection strength to corn of interior sacculus study than ever pure sandy soil S formulation medias be respectively increased 20/g, 10/
g。
5) microbial inoculum small volume, quality transport storage easily.
Description of the drawings
Fig. 1 is the determination for infecting grade.
Fig. 2 is the mould clump branch of Moses's pipe handle capsule and vesicle structure in sandy soil matrix maize root system.
Fig. 3 is the mould clump branch of Moses's pipe handle capsule and vesicle structure in weathering matrix of coal maize root system.
Fig. 4 is the vesicle structure and spore for connecing Glomus intraradices in bacterium maize root system.
Fig. 5 is nonvaccinated maize root system.
Fig. 6 is indoor three kinds of upgrowth situations processed after corn 2 first quarter moons of culture, and left figure is that the difference of Z formulas is connect at bacterium
Reason, right figure are that the difference of W formulas connects bacterium process, it can be seen that connect bacterium apparently higher than control treatment, and Gm-Z growing ways are better than Gi-
Z, Gi-S, and Gi-W growing ways are better than Gm-W, Gi-S.
Specific embodiment
Experimental technique used in following embodiments if no special instructions, is conventional method.
In following embodiments, material used, reagent etc., if no special instructions, commercially obtain.
Embodiment 1, AM fungal inoculants expand numerous method and set up
First, AM fungies are expanded numerous method and set up
The expanding propagation method flow process of AM fungal inoculants：Substrate composition corn germination microbial inoculum is inoculated with seedling management
1st, culture matrix proportioning：Sandy soil, vermiculite, perlite are crossed into high pressure steam sterilization (121 DEG C, 1h) after 2mm sieves, with thin
Film dries standby after covering 3 the Heavenly Stems of drying；Sandy soil, weathered coal cross 2mm sieves, the vermiculite of appropriate 1-2mm particle diameters, perlite (this
In test, sandy soil are bought in northern sandy beach, and weathered coal picks up from Inner Mongol coal field, and vermiculite, perlite are in buying in matrix factory)
Standby (being volume ratio in bracket) is mixed according to following proportioning：
S formula==pure sandy soil；
Z formula=sandy soil (2): vermiculite (1): perlite (1)；
W formula==sandy soil (1): vermiculite (1): perlite (1)：Weathered coal (1).
2nd, host plant presprouting of seeds
Select full seed, without damaged, the cultivation corn seed without insect bite and clover seed, first use volume hundred respectively
Content is divided to be 10%H2O2The aqueous solution sterilizes 10min, then with aseptic water washing 5 times, then is placed in the culture dish of moistening and places,
Temperature is dark vernalization 3d in 28 DEG C of constant incubators, obtains after vernalization clover seed after corn seed and vernalization.
3rd, inoculation is expanded numerous
The sterilization of container：75% ethanol solution wipes the plastic flowerpot of 18cm × 24cm.
Inoculation：First fill above-mentioned the 1 of 2/3 3 kinds of culture matrixes in each flowerpot respectively, excuted a law using bar and add 50g clumps
(with pure sandy soil as culture matrix, corn is host plant to mycorrhizal fungi microbial inoculum, cultivates 100 days, collect after inoculation clump branch bacterium
The bush mycorrhizal fungi preparation that host plant root system and its Rhizosphere Soil are obtained after being cultivated), then 3 vernalization are spread respectively per basin
Clover seed after corn seed or 20-25 grain vernalization, with remaining matrix filling in basin, waters to maximum water-holding capacity afterwards；Culture.
S, Z, W formula irrigation amount is respectively 25%, 50% and the 70% of matrix volume.
Irrigation amount is adjusted according to weather condition, room temperature height, evaporation speed moisture content of weighing or determine, 300-
500ml is advisable；Temperature adjusting is between 22-30 DEG C.Suitably prevented and treated depending on pest and disease damage situation.
Emerge after 4d, 20 plants of 1 plant of corn or clover, configuration Hoagland nutrient solution mother liquors (being shown in Table 1) are colonized per basin；
Seedling Stage, pours the Hoagland nutrient solutions of 1/10 intensity after emerging one week；
After seedling (after five leaves), a 1/5 intensity nutrient solution is poured weekly；
Harvest time (after seven leaves), after no longer pouring nutrient solution at least 2 months, above in relation to sandy soil culture matrix.
Table 1 is Hoagland compoundings
Note：One hurdle of the rightmost side represents the mother liquor ml prepared needed for 1L nutrient solutions
The root system and culture matrix of the host plant after cultivating after culture 3 months, are collected, that is, obtains different substrates acquisition
AM fungal inoculants, realize expand it is numerous.
Fig. 2 is the mould clump branch of Moses's pipe handle capsule and vesicle structure in sandy soil matrix maize root system.Fig. 3 is that weathering matrix of coal is beautiful
The mould clump branch of Moses's pipe handle capsule and vesicle structure in rice root system.Fig. 4 is the vesicle structure for connecing Glomus intraradices in bacterium maize root system
And spore.
2nd, AM fungal inoculants quality analysis
AM fungal infections status investigation in AM fungal inoculants is carried out using Trayman Blue colouring methods, arbuscular mycorrhiza is used
(1986) Trourelot.etc determines infection strength, determines AM fungies with wet screening decantation the appraisal procedure of fungal infection
Spore amount, carries out the quality evaluation of microbial inoculum according to this.
1st, trypan blue improvement Determination Staining infection strength
1) trypan blue improvement decoration method dyeing
The capillary root of the AM fungal inoculant root systems that above-mentioned is obtained, is cut into the long root segments of l cm and is placed in culture dish after cleaning
In, weight/mass percentage composition 10%KOH aqueous solution soakings water-bath 60min in 90 DEG C of water-baths is added, alkali lye after cooling, is discarded, is used
Clear water is washed away 3-5 time；Add and acid solution is gone after 5min being acidified under weight/mass percentage composition 2%HCl room temperatures, add 0.05% by sharp benzene
Blue dyeing liquor water-bath dyeing 30min in 90 DEG C, after discarding dye liquor cooling, uses 1:1:It is many that the immersion of 1 lactic acid glycerol liquor removes root sample
Remaining dyestuff, makes mycelia, vesicle and the clump branch of mycorhiza keep colored state, film-making microscopy.
2) calculate infection strength and infect grade (Trouvelot A, 1986)
A. place in the AM fungal inoculants (host is corn) for above-mentioned being obtained and AM fungal inoculants (host is clover)
15 root segments of main root system of plant are fixed on 1 slide, 2 slice, thin pieces of plant fibrous root root segment system after 30 dyeing；
B. these root segments are examined under a microscope, determines that classification (refers to https according to the sorting technique of Fig. 1://
Www2.dijon.inra.fr/mychintec/Mycocalc-prg/download.html), this classification is included to each root
The rapid evaluation of section mycorhiza infection strength level and clump branch abundance.
C. correlation is calculated according to following correlation formula by substitute into observation in computer software " Mycocale "
Mycorhiza degree of infecting parameter, expands numerous effect to microbial inoculum and evaluates：%M, and %A (Trouvelot et.al 1986；Following inspection
Index and computing formula are surveyed known in existing, https is referred to://www2.dijon.inra.fr/mychintec/Mycocalc-
prg/download.h)
Mycorhiza infection strength (%M) in root system=(more than 90% root segment number+70* of 95* infection strengths infect 50% to
Root segment number+5* infection strengths more than 1% root segment number below 10% of 90% root segment number+30* infection strengths 10% to 50%+invade
Dye intensity root segment number below 1%)/total root segment number * 100
It is total root segment numbers of mycorhiza infection strength (%m)=M* in root segment/have root segment number with respect to mycorhiza intensity
(with respect to clump branch rate) root segment clump branch rate (%a)=(100*mA3+50mA2+10mA1)/100, wherein mA3, mA2,
MA1 is A3 respectively, the mycorhiza infection strength corresponding to A2, A1, A3, A2, and A1 is shown in Fig. 1.
(absolute clump branch rate) root system clump branch rate (A%)=a* (M/100)
2nd, wet screening decantation determines the spore amount of AM fungies
The dry ground earth near AM fungal inoculant root systems that above-mentioned one is obtained weighs g, is placed in container the 20- that is soaked in water
30min, makes looseness of soil, from a set of with the soil sieve that aperture is 0.5-0.034mm, overlaps successively, one object of bottom
(such as wooden unit) is cushioned, makes compass screen surface slightly incline.The aqueous solution of immersion is stirred with glass bar, precipitates big debris within parked several seconds, will
Suspension is poured on the maximum soil sieve in most last layer aperture slowly.When toppling over, it is poured on a point of compass screen surface, with clear water successively gently
Flushing rests on the outsifting on compass screen surface, and the culture that the outsifting on compass screen surface is gently flushed to a cleaning is will stay on wash bottle
Inside ware, then filtrate is passed through into dusting cover and is rinsed with water, the different-diameter in the outsifting for sweeping away just containing AM mycorrhizal fungis
Spore, will containing undersize culture dish be placed under binocular body formula disecting microscope observation (Gerdemann J W,
Nicolson T H, 1963.).
The inoculation of embodiment 2, AM fungies from numerous microbial inoculum
First, AM fungies inoculation
1st, culture matrix proportioning：Same as Example 1, matrix includes S proportionings, Z proportionings and W proportionings；
2nd, host plant presprouting of seeds：It is same as Example 1；Host plant is corn, obtains corn seed after vernalization.
3rd, it is inoculated with：It is same as Example 1；
The sterilization of container：75% ethanol solution wipes the plastic flowerpot of 18cm × 24cm.
Inoculation：First fill above-mentioned the 1 of 2/3 3 kinds of culture matrixes in each flowerpot respectively, excuted a law under addition 50g using bar
Face bush mycorrhizal fungi preparation, then spreads corn seed after 3 vernalization respectively per basin, with remaining matrix filling in basin, water to
Maximum water-holding capacity, culture 3-5 month.
Bush mycorrhizal fungi preparation is respectively Moses's pipe handle capsule mould agent and Glomus intraradices microbial inoculum；
For trying microbial inoculum：Above-mentioned Moses's pipe handle capsule mould agent is that corn is host plant as culture matrix with pure sandy soil,
Inoculation Moses's pipe handle capsule it is mould (Glomus mosseae, write a Chinese character in simplified form Gm, be documented in " Cui Weidong, dragon a surname's Qi, Hou Xin are strong etc.
Verticillium wilt opportunistic pathogen is coerced on arbuscular mycorrhiza cotton seedling root protective enzyme and Ultrastructural impact. Xinjiang Agricultural Sciences,
2009,46(6):1235-1244 " one is literary) cultivate 3 months afterwards, after collecting culture, the root system and culture matrix of host plant is obtained
Moses's pipe handle capsule mould agent.
Glomus intraradices microbial inoculum is that corn is host plant as culture matrix with pure sandy soil, is inoculated with Glomus intraradices
(Glomus intraradices, write a Chinese character in simplified form Gi, are documented in Kuhn G, Hijri M, Sanders IR, Evidence for the
evolution of multiple genomes in arbuscular mycorrhizal fungi.Nature,2001,
414:745～748) cultivate 3 months afterwards, the Glomus intraradices bacterium that the root system and culture matrix of host plant is obtained after collecting culture
Agent.
After cultivating 3 months in 3 kinds of culture matrixes, after collection culture, corn infects root system and the rhizosphere with brood body
Soil, that is, obtain expand it is numerous after AM fungal inoculant Gm-Z (bacterial classification is Gm, and matrix is Z), Gm-W (bacterial classification is Gm, and matrix is W), Gm-S
(bacterial classification is for (bacterial classification is Gm, and matrix is S), Gi-Z (bacterial classification is Gi, and matrix is Z), Gi-W (bacterial classification is Gi, and matrix is W), Gi-S
Gi, matrix are S).
2nd, microbial inoculum quality analysis
1st, the leaf color value determined using chlorophyll meter
(production of Zhejiang Top) is determined using SPAD-502 chlorophyll meters is carried out to the leaf color value of each plant before results
Determine, obtain data as shown in table 2, the leaf color value of Gm-W be significantly increased than Gm-Z phases, leaf color value and the Gm-W phases of Gi-Z
Than also increasing, but difference is not notable.
The leaf color value of 2 corn of table
2nd, trypan blue improvement Determination Staining infects degree
Trypan blue improvement Determination Staining the inventive method expand numerous expansion for obtaining it is numerous after AM fungal inoculant Gm-Z, Gm-W,
The infection strength of Gm-S, Gi-Z, Gi-W, Gi-S, method are same as Example 1.
As a result it is as shown in table 3, it can be seen that Glomus intraradices, are rubbed to the mycorhiza infection strength of corn more than 72%
, more than 91%, matrix Z Moses's pipe handle capsule is mould, Glomus intraradices are to corn for the mould mycorhiza infection strength to corn of western pipe handle capsule
Mycorhiza infection strength study pure sandy soil than ever be respectively increased 6.64%, 15.51%, matrix W Moses's pipe handle capsule is mould, root
What the mould mycorhiza infection strength to corn of interior sacculus studied pure sandy soil than ever has been respectively increased 6.72%, 8.19%.
Table 3 corn infects degree
3rd, wet screening decantation determines the spore amount of AM fungies
Expand numerous corn AM fungal spores density result as shown in table 4,3 kinds of matrix are mould to Moses's pipe handle capsule, sacculus in root
Fungal spore density in mould 2 kinds of AM fungal culture matrix is more than 70/g, compared with pure sandy soil spore density, Gm-W, Gi-W
The mycorhiza infection strength of corn is studied than ever pure sandy soil be respectively increased 20/g, 4/g, matrix Gm-Z, Gi-Z pair
What the mycorhiza infection strength of corn studied pure sandy soil than ever has been respectively increased 32/g, 10/g (table 3).
Table 4 expand the spore density evaluation of numerous AM fungal inoculants/
Prove with reference to Fig. 5,6 and table 1-4 tables, the W formula (weathered coals (2) for filtering out:Sandy soil (1):Vermiculite (1):Perlite
(1)) be Gm microbial inoculums the numerous matrix of most preferably expansion, Z formula (sandy soil (2):Vermiculite (1):Perlite (1)) be Gi microbial inoculums most preferably expansion it is numerous
Matrix, with this understanding, infects degree, spore density and leaf color value optimal.
Claims (9)
1. a kind of preparation method of AM fungal inoculants, comprises the steps A：The host plant of AM fungies is being applied with into AM fungies
Culture matrix in cultivate, collect culture after the host plant root system and culture matrix, that is, obtain AM fungal inoculants；
The culture matrix for it is following 1) or 2)：
1) culture matrix shown in is made up of sandy soil, vermiculite and perlite；
2) culture matrix shown in is made up of sandy soil, vermiculite, perlite and weathered coal.
2. method according to claim 1, it is characterised in that：
1), in the culture matrix shown in, the sandy soil, the vermiculite and the perlitic volume ratio are 2-3:1:1；
2), in the culture matrix shown in, the volume ratio of the sandy soil, the vermiculite, the perlite and the weathered coal is 1-2:
1:1:1。
3. method according to claim 1 and 2, it is characterised in that：
After the collection culture, the root system of host plant and the time of culture matrix are to form host plant after per gram of culture
Root system and the AM fungal spore quantity of culture matrix when reach the amount of 40-50；
Or the incubation time is no less than 3 months；
Or the incubation time is 3-5 month.
4. according to arbitrary described method in claim 1-3, it is characterised in that：
In methods described, also include the step of host plant seed is carried out into vernalization before culture.
5. method according to claim 4, it is characterised in that：The condition of the vernalization is 28-30 DEG C of light culture 3 days.
6. according to arbitrary described method in claim 1-5, it is characterised in that：
The host plant is can be with the plant of AM mycosymbiosises；
Or, described can be corn, Chinese sorghum, clover with the plant of AM mycosymbiosises.
7. according to arbitrary described method in claim 1-6, it is characterised in that：The AM fungies be Moses's pipe handle capsule mould or
Glomus intraradices bacterium.
8. according to arbitrary described method in claim 1-7, it is characterised in that：
The corresponding culture matrix of Moses's pipe handle capsule mould for it is described 2) shown in culture matrix；
The corresponding culture matrix of the Glomus intraradices bacterium for it is described 1) shown in culture matrix.
9. a kind of AM fungies expanding propagation method, including step A in claim 1-8 in arbitrary methods described, realizes that fungi is expanded numerous.
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| CN110066738A (en) * | 2019-04-02 | 2019-07-30 | 亿利资源集团有限公司 | A kind of AM fungal inoculant and its application method |
| CN111165266A (en) * | 2020-01-08 | 2020-05-19 | 华南农业大学 | Method for continuously expanding propagation of arbuscular mycorrhizal fungal spores through compartment culture |
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| CN112189525A (en) * | 2020-10-09 | 2021-01-08 | 四川农业大学 | Method for promoting leguminous plant growth by using arbuscular mycorrhizal fungi and rhizobia |
| CN113024313A (en) * | 2021-04-28 | 2021-06-25 | 上海乾界生物科技有限公司 | Mycorrhizal fungi agent and preparation method thereof |
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| CN114175996A (en) * | 2021-12-08 | 2022-03-15 | 青岛农业大学 | Propagation method of arbuscular mycorrhizal fungi |
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