CN103314676A - Method for carrying out industrialized production on arbuscular mycorrhizal fungi agent - Google Patents
Method for carrying out industrialized production on arbuscular mycorrhizal fungi agent Download PDFInfo
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Abstract
The invention discloses a method for carrying out industrialized production on an arbuscular mycorrhizal fungi agent, and relates to a production method of arbuscular mycorrhizal fungi. The method comprises the following steps: 1. preparing and treating the arbuscular mycorrhizal fungi agent; 2. carrying out seed treatment on hosts; 3. carrying out breeding; and 4. managing. The method disclosed by the invention has the advantages that the implementation of production just needs eight weeks, the number of effective spores of arbuscular mycorrhizal fungi in each gram of the obtained arbuscular mycorrhizal fungi agent is 120-150, the production cycle is short, and the yield is high. The method disclosed by the invention can be applied to the field of agriculture, gardens, and flowers.
Description
Technical field
The present invention relates to a kind of production method of bush mycorrhizal fungi preparation.
Background technology
Mycorhiza (mycorrhiza) is a kind of general plant symbiosis phenomenon of occurring in nature, and it is mycorrhizal fungi in the soil and a kind of symbiont of higher plant root growth.Arbuscular mycorrhiza (Arbuscular mycorrhiza, AM) is a kind of modal endotrophic mycorrhiza, is the symbiont that a kind of arbuscular mycorrhizal fungi in the soil and root system of plant form.Arbuscular mycorrhizal fungi is widely distributed at occurring in nature, on the known world about 90% flowering plant and pteridophyte and bryophyte can with the arbuscular mycorrhizal fungi symbiosis.
Arbuscular mycorrhizal fungi plays an important role to the balance of plant nutrient circulation and effective utilization of moisture, but Promoting plant growth improves the plant transplantation survival rate, and can improve the resistance of plant.And many beneficial effects that the AM fungi produces growth and development of plants particularly improve aspect plant phosphorus element, nitrogen and the mineral nutrition the AM fungi.There are some researches show, Inoculation of Arbuscular Mycorrhizal Fungi under the certain condition can promote plant to absorption and the utilization of Soil Phosphorus, zinc, copper, and absorbing also to nitrogen, potassium, magnesium, sulphur, manganese etc., tool has certain effect.Therefore, arbuscular mycorrhizal fungi is with a wide range of applications in agriculture and forestry production and afforestation field.
The average usage amount of China's nitrogenous fertilizer is nearly 3 times, more than 8 times of Australia of the U.S., and this has caused the waste of a large amount of non-renewable energy resources, and the farmland soil property degenerates, and fertility descends, and crops quality reduces, and the environmental situations such as food and underground water also are on the rise.Therefore, significant to agricultural, resource, environment of China development bush mycorrhizal fungi preparation.
But arbuscular mycorrhizal fungi is the strict symbiosis fungi of a class, and its existence strictly depends on the live body higher plant, so it is larger to cultivate on a large scale and produce the bush mycorrhizal fungi preparation difficulty.The method that obtains at present arbuscular mycorrhizal fungi in the laboratory has a lot, as: the aseptic dual cultivation of medium culture method, static nutrient solution cultivation method, the nutrient solution cultivation method that flows, spray liquid training method, AM fungi and Vitro Plant root organ and bead locellus cultivation etc.; But cheap, the large-scale method that can be used as bush mycorrhizal fungi preparation production only has potted plant cultivation and land for growing field crops cultivation., potted plant cultivation and land for growing field crops cultivation the production cycle long (4～5 months) arranged, be subjected to that such environmental effects is large, microbial inoculum is produced in limited quantities, sporogenesis body quantity few (80～100/g microbial inoculum), quality is unstable and Cultivate administration is loaded down with trivial details problem.
Summary of the invention
There is production cycle length in cheap, the large-scale method that the present invention will solve existing bush mycorrhizal fungi preparation production, is subjected to the problem that such environmental effects is large, microbial inoculum is produced in limited quantities, sporogenesis body quantity is few, quality is unstable and Cultivate administration is loaded down with trivial details, and the method that bush mycorrhizal fungi preparation is produced in a kind of batch production that provides.
Batch production of the present invention is produced the method for bush mycorrhizal fungi preparation and is carried out according to the following steps:
One, soil, sand and vermiculite are mixed into matrix by the volume ratio of 5:2:3,50 times of the formalin dilute with waters that will contain again 40% formaldehyde evenly are sprayed onto on the matrix afterwards, then with Polypropylence Sheet sealing 1 round the clock, spread out afterwards, insolation is 4～6 days in the sun, namely obtains the bush mycorrhizal fungi preparation culture matrix;
Two, Chinese sorghum, clover or trefoil seed are washed by rubbing with the hands 2～3 times with 20～30 ℃ of warm water, be that 0.3% liquor potassic permanganate soaked 3～4 hours with concentration again, then eluriate clean with clear water, afterwards again with 20～30 ℃ Seed soaking 24 hours and eluriate 2～3 times, host's seed treatment is finished in vernalization;
Three, the bush mycorrhizal fungi preparation culture matrix being put into is the culture pond of common brick for cement concrete, bottom all around, then treated host's seed and arbuscular mycorrhizal fungi are imposed on bush mycorrhizal fungi preparation culture matrix surface jointly, spray concentration is 5 of 150nmol/L again, 7,4'-trihydroxyflavone solution, cladding thickness is the bush mycorrhizal fungi preparation culture matrix of 0.4～0.7cm more afterwards; Wherein, the consumption of Apigenin solution is 250mL/ square metre; The culture pond degree of depth is 0.25～0.35 meter;
Four, conventional field management, be cultured to the cauline leaf part of removing host plant the 7th week, continue to cultivate to the main root of removing again host plant in the bush mycorrhizal fungi preparation culture matrix the 8th week, adventive root and bush mycorrhizal fungi preparation culture matrix mixing, granulation, namely obtain bush mycorrhizal fungi preparation.
The inventive method has to produce only needed for 8 weeks, and effective spore quantity of arbuscular mycorrhizal fungi is 120～150 in the every gram bush mycorrhizal fungi preparation that obtains, have advantages of with short production cycle, output is high.
All culture matrixes of the inventive method are cheap, and simple to operate, are fit to the production of extensive bush mycorrhizal fungi preparation.Because the inventive method is by the processing to bush mycorrhizal fungi preparation culture matrix and host's seed, and the designing and arranging of culture pond is except the interference of other factors to the arbuscular mycorrhizal fungi breeding, promote the output of arbuscular mycorrhizal fungi, and significantly improved the stability of product.And conventional method is adopted in field management of the present invention, and it is time saving and energy saving to have advantages of.
Description of drawings
Fig. 1 is that Radix Sorghum vulgare Pers is subjected to Glomus mosseae to infect microscopic examination figure among the embodiment 1.
Embodiment
Technical solution of the present invention is not limited to following cited embodiment, also comprises any combination between each embodiment.
Embodiment one: the present embodiment batch production is produced the method for bush mycorrhizal fungi preparation and is carried out according to the following steps:
One, soil, sand and vermiculite are mixed into matrix by the volume ratio of 5:2:3,50 times of the formalin dilute with waters that will contain again 40% formaldehyde evenly are sprayed onto on the matrix afterwards, then with Polypropylence Sheet sealing 1 round the clock, spread out afterwards, insolation is 4～6 days in the sun, namely obtains the bush mycorrhizal fungi preparation culture matrix;
Two, Chinese sorghum, clover or trefoil seed are washed by rubbing with the hands 2～3 times with 20～30 ℃ of warm water, be that 0.3% liquor potassic permanganate soaked 3～4 hours with concentration again, then eluriate clean with clear water, afterwards again with 20～30 ℃ Seed soaking 24 hours and eluriate 2～3 times, host's seed treatment is finished in vernalization;
Three, the bush mycorrhizal fungi preparation culture matrix being put into is the culture pond of common brick for cement concrete, bottom all around, then treated host's seed and arbuscular mycorrhizal fungi are imposed on bush mycorrhizal fungi preparation culture matrix surface jointly, spray concentration is 5 of 150nmol/L again, 7,4'-trihydroxyflavone solution, cladding thickness is the bush mycorrhizal fungi preparation culture matrix of 0.4～0.7cm more afterwards; Wherein, the consumption of Apigenin solution is 250mL/ square metre; The culture pond degree of depth is 0.25～0.35 meter;
Four, conventional field management, be cultured to the cauline leaf part of removing host plant the 7th week, continue to cultivate to the main root of removing again host plant in the bush mycorrhizal fungi preparation culture matrix the 8th week, adventive root and bush mycorrhizal fungi preparation culture matrix mixing, granulation, namely obtain bush mycorrhizal fungi preparation.
Plant host is selected in the present embodiment is seed after the vernalization, and the seed that has germinateed can directly be infected by arbuscular mycorrhizal fungi, can more early set up syntaxial system with plant host, improves infection rate; The root synchronous growth of arbuscular mycorrhizal fungi and plant host can increase more effectively and rapidly infection rate, increase mycelia breeding amount and spore quantity.Because the foundation that syntaxial system just begins when the seed germination state, plant host is also benefited from it, so the root growth of plant host and growth is faster, better, what distribute in the bush mycorrhizal fungi preparation culture matrix is wider; Meanwhile, arbuscular mycorrhizal fungi has also obtained a large amount of propagation, and extends to the everywhere of culture matrix along with the plant host Root growth.
Although, 5,7,4'-trihydroxyflavone belongs to the flavonoids material and can be used as arbuscular mycorrhizal fungi mycorhiza signal recognition material, has obtained the approval of scientific circles, but because 5,7,4'-trihydroxyflavone is water insoluble, and 5, content and the concentration thereof of 7,4'-trihydroxyflavone in soil there is no clear and definite research conclusion to the growth effect of arbuscular mycorrhizal fungi.Therefore, Apigenin can't be actually used in the production of arbuscular mycorrhizal fungi all the time.Present embodiment has been determined using dosage and the using method of Apigenin, and has obtained significantly technique effect.
Present embodiment is selected with soil, sand and vermiculite as the bush mycorrhizal fungi preparation culture matrix, has convenience of drawing materials, cheap, the plurality of advantages such as good water permeability, and not only be conducive to the growth of plant host under this ratio condition, also meet the growing environment of arbuscular mycorrhizal fungi.
The problems such as the employing conventional method is sterilized to matrix, has energy consumption high, and cost is large, and labour intensity is large, effect is undesirable.Present embodiment selects chemistry (chemicals sprinkling) and physics (sunlight insolation) dual mode to carry out disinfection, wherein spray and contain the reinforcement that utilizes its steam matrix to be carried out 24 hours behind the formalin dilution of formaldehyde, sterilization effect is good, by the sunlight insolation, not only can utilize ultraviolet sterilization, degerming in the sunlight, can also effectively remove residual formaldehyde and formalin.The sterilization method that present embodiment is selected can keep the mesotrophic composition of matrix chemical change not occur.
Present embodiment limits the degree of depth of culture pond, this depth bounds can guarantee the needs of host's plant root growth and the demand of nutrient, can within campaign, make again the host's plant root with the symbiosis arbuscular mycorrhizal fungi be full of the bush mycorrhizal fungi preparation culture matrix, make distribution and the uniform content of arbuscular mycorrhizal fungi, thereby guaranteed the uniformity of the interior arbuscular mycorrhizal fungi mycelium of bush mycorrhizal fungi preparation and spore.
Be cement concrete, bottom place mat common brick around the present embodiment culture pond, the interference that can avoid like this extraneous factor that bush mycorrhizal fungi preparation is produced can be discharged unnecessary moisture again, has advantages of gas permeability, good water permeability.
Present embodiment is cultured to the cauline leaf part of removing host plant the 7th week, is in order to stop the continued growth of root system, to promote the speed of spore maturation, and effectively increase spore quantity.
Continuing to cultivate to the main root of removing again host plant in the bush mycorrhizal fungi preparation culture matrix the 8th week, is because do not contain the subsidiary body of mycelia and the fungi thereof of arbuscular mycorrhizal fungi in the main root, also can affect simultaneously granulation and the processing of microbial inoculum.
Cladding thickness is the bush mycorrhizal fungi preparation culture matrix of 0.4～0.7cm on host's seed of vernalization, and this thickness can guarantee that host plant germinates, emerge fast in the process of growth and take root solidly, also is conducive to the mycorrhizal fungal spore fast-germination simultaneously.
Present embodiment is cultivated the 8th all Mycorrhizal Infection Incidences and is almost reached 100%, and effective spore quantity of arbuscular mycorrhizal fungi is all above 120 in every gram bush mycorrhizal fungi preparation.
Embodiment two: the difference of present embodiment and embodiment one is: every cubic metre of matrix is sprayed with the formalin dilution that 500mL contains formaldehyde in the step 1.Other step and parameter are identical with embodiment one.
Embodiment three: present embodiment and embodiment one or twos' difference is: in the step 2 length of culture pond be 5 meters, wide be 2 meters.Other step and parameter are identical with embodiment one or two.
Embodiment four: present embodiment and embodiment one, two or three difference are: the arbuscular mycorrhizal fungi in the step 3 is Glomus intraradices (Glomus intraradices), Scotland sacculus mould (Glomus caledonium), earth's surface sacculus mould (Glomus versiforme) or Glomus mosseae (Glomus mosseae).Other step and parameter and embodiment one, two or three identical.
Embodiment five: the difference of one of present embodiment and embodiment one to four is: every square metre on bush mycorrhizal fungi preparation culture matrix surface adds 25000～30000 spores of arbuscular mycorrhizal fungi in the step 3.Other step and parameter are identical with one of embodiment one to four.
Embodiment 1
One, soil, sand and vermiculite are mixed into matrix by the volume ratio of 5:2:3,50 times of the formalin dilute with waters that will contain again 40% formaldehyde evenly are sprayed onto on the matrix afterwards, then with Polypropylence Sheet sealing 1 round the clock, spread out afterwards, insolation is 5 days in the sun, namely obtains the bush mycorrhizal fungi preparation culture matrix;
Two, the seed of Chinese sorghum is washed by rubbing with the hands 3 times with 20 ℃ of warm water, is that 0.3% liquor potassic permanganate soaked 3 hours with concentration again, then eluriates totally with clear water, and again with 30 ℃ Seed soaking 24 hours and eluriate 2 times, host's seed treatment is finished in vernalization afterwards;
Three, the bush mycorrhizal fungi preparation culture matrix being put into is the culture pond of common brick for cement concrete, bottom all around, then treated host's seed and Glomus mosseae are imposed on bush mycorrhizal fungi preparation culture matrix surface jointly, spray concentration is 5 of 150nmol/L again, 7,4'-trihydroxyflavone solution, cladding thickness is the bush mycorrhizal fungi preparation culture matrix of 0.7cm more afterwards; Wherein, the consumption of Apigenin solution is 250mL/ square metre; The culture pond degree of depth is 0.3 meter;
Four, conventional field management, be cultured to the cauline leaf part of removing host plant the 7th week, continue to cultivate to the main root of removing again host plant in the bush mycorrhizal fungi preparation culture matrix the 8th week, adventive root and bush mycorrhizal fungi preparation culture matrix mixing, granulation, namely obtain bush mycorrhizal fungi preparation; Wherein, every cubic metre of matrix is sprayed with the formalin dilution that 500mL contains formaldehyde in the step 1; In the step 2 length of culture pond be 5 meters, wide be 2 meters; Every square metre on bush mycorrhizal fungi preparation culture matrix surface adds 25000 spores of Glomus mosseae in the step 3.
Adopt the acid fuchsin colouring method of Phillip and Hayman to measure Mycorrhizal Infection Incidence to Sorghum roots weekly, when the 8th week, adopt wet screening to microbial inoculum in fungal spore detect, testing result is as shown in table 1.
Table 1
| ? | The 2nd week | The 4th week | The 6th week | The 8th week |
| Mycorrhizal Infection Incidence (%) | 52.6±1.5 | 96.8±1.2 | 98.4±0.8 | 99.9±0.1 |
| Microbial inoculum miospore content (individual/g) | ? | ? | ? | 135±6 |
By to the statistical analysis of many groups sampled data, sorghum seedling is when growing into for the 2nd week, and Mycorrhizal Infection Incidence has reached more than 50%, and Mycorrhizal Infection Incidence is up to more than 95% when the 4th week.Particularly the spore content of Glomus mosseae shows that up to 135 spore content is abundant in the 8th when week every gram culture matrix.
Embodiment 2
One, soil, sand and vermiculite are mixed into matrix by the volume ratio of 5:2:3,50 times of the formalin dilute with waters that will contain again 40% formaldehyde evenly are sprayed onto on the matrix afterwards, then with Polypropylence Sheet sealing 1 round the clock, spread out afterwards, insolation is 6 days in the sun, namely obtains the bush mycorrhizal fungi preparation culture matrix;
Two, the seed of clover is washed by rubbing with the hands 3 times with 25 ℃ of warm water, is that 0.3% liquor potassic permanganate soaked 3.5 hours with concentration again, then eluriates totally with clear water, and again with 25 ℃ Seed soaking 24 hours and eluriate 3 times, host's seed treatment is finished in vernalization afterwards;
Three, the bush mycorrhizal fungi preparation culture matrix being put into is the culture pond of common brick for cement concrete, bottom all around, then treated host's seed and arbuscular mycorrhizal fungi are imposed on bush mycorrhizal fungi preparation culture matrix surface jointly, spray concentration is 5 of 150nmol/L again, 7,4'-trihydroxyflavone solution, cladding thickness is the bush mycorrhizal fungi preparation culture matrix of 0.5cm more afterwards; Wherein, the consumption of Apigenin solution is 250mL/ square metre; The culture pond degree of depth is 0.3 meter;
Four, conventional field management, be cultured to the cauline leaf part of removing host plant the 7th week, continue to cultivate to the main root of removing again host plant in the bush mycorrhizal fungi preparation culture matrix the 8th week, adventive root and bush mycorrhizal fungi preparation culture matrix mixing, granulation, namely obtain bush mycorrhizal fungi preparation; Wherein, every cubic metre of matrix is sprayed with the formalin dilution that 500mL contains formaldehyde in the step 1; In the step 2 length of culture pond be 5 meters, wide be 2 meters; Every square metre on bush mycorrhizal fungi preparation culture matrix surface adds 27000 spores of Glomus intraradices in the step 3.
Adopt the acid fuchsin colouring method of Phillip and Hayman to measure Mycorrhizal Infection Incidence to the clover root system weekly, when the 8th week, adopt wet screening to microbial inoculum in fungal spore detect, testing result is as shown in table 2.
Table 2
| ? | The 2nd week | The 4th week | The 6th week | The 8th week |
| Mycorrhizal Infection Incidence (%) | 51.9±1.3 | 96.2±1.1 | 98.3±0.9 | 99.8±0.1 |
| Microbial inoculum miospore content (individual/g) | ? | ? | ? | 134±5 |
By to the statistical analysis of many groups sampled data, seedling of alfalfa is when growing into for the 2nd week, and Mycorrhizal Infection Incidence has reached more than 50%, and Mycorrhizal Infection Incidence is up to more than 95% when the 4th week.Particularly the spore content of Glomus intraradices shows that up to 134 spore content is abundant in the 8th when week every gram culture matrix.
Embodiment 3
One, soil, sand and vermiculite are mixed into matrix by the volume ratio of 5:2:3,50 times of the formalin dilute with waters that will contain again 40% formaldehyde evenly are sprayed onto on the matrix afterwards, then with Polypropylence Sheet sealing 1 round the clock, spread out afterwards, insolation is 4 days in the sun, namely obtains the bush mycorrhizal fungi preparation culture matrix;
Two, trefoil seed is washed by rubbing with the hands 3 times with 20 ℃ of warm water, is that 0.3% liquor potassic permanganate soaked 4 hours with concentration again, then eluriates totally with clear water, and again with 30 ℃ Seed soaking 24 hours and eluriate 3 times, host's seed treatment is finished in vernalization afterwards;
Three, the bush mycorrhizal fungi preparation culture matrix being put into is the culture pond of common brick for cement concrete, bottom all around, then treated host's seed and arbuscular mycorrhizal fungi are imposed on bush mycorrhizal fungi preparation culture matrix surface jointly, spray concentration is 5 of 150nmol/L again, 7,4'-trihydroxyflavone solution, cladding thickness is the bush mycorrhizal fungi preparation culture matrix of 0.6cm more afterwards; Wherein, the consumption of Apigenin solution is 250mL/ square metre; The culture pond degree of depth is 0.35 meter;
Four, conventional field management, be cultured to the cauline leaf part of removing host plant the 7th week, continue to cultivate to the main root of removing again host plant in the bush mycorrhizal fungi preparation culture matrix the 8th week, adventive root and bush mycorrhizal fungi preparation culture matrix mixing, granulation, namely obtain bush mycorrhizal fungi preparation; Wherein, every cubic metre of matrix is sprayed with the formalin dilution that 500mL contains formaldehyde in the step 1; In the step 2 length of culture pond be 5 meters, wide be 2 meters; Every square metre on bush mycorrhizal fungi preparation culture matrix surface adds mould 30000 spores of Scotland sacculus in the step 3.
Adopt the acid fuchsin colouring method of Phillip and Hayman to measure Mycorrhizal Infection Incidence to the clover root system weekly, when the 8th week, adopt wet screening to microbial inoculum in fungal spore detect, testing result is as shown in table 3.
Table 3
| ? | The 2nd week | The 4th week | The 6th week | The 8th week |
| Mycorrhizal Infection Incidence (%) | 53.2±1.7 | 97.0±1.0 | 98.3±0.5 | 99.9±0.1 |
| Microbial inoculum miospore content (individual/g) | ? | ? | ? | 134±6 |
By to the statistical analysis of many groups sampled data, the clover seedling is when growing into for the 2nd week, and Mycorrhizal Infection Incidence has reached more than 50%, and Mycorrhizal Infection Incidence is up to more than 95% when the 4th week.Particularly sacculus mould spore content in Scotland shows that up to 134 spore content is abundant in the 8th when week every gram culture matrix.
Embodiment 4
One, soil, sand and vermiculite are mixed into matrix by the volume ratio of 5:2:3,50 times of the formalin dilute with waters that will contain again 40% formaldehyde evenly are sprayed onto on the matrix afterwards, then with Polypropylence Sheet sealing 1 round the clock, spread out afterwards, insolation is 5 days in the sun, namely obtains the bush mycorrhizal fungi preparation culture matrix;
Two, the seed of Chinese sorghum is washed by rubbing with the hands 3 times with 25 ℃ of warm water, is that 0.3% liquor potassic permanganate soaked 4 hours with concentration again, then eluriates totally with clear water, and again with 28 ℃ Seed soaking 24 hours and eluriate 2 times, host's seed treatment is finished in vernalization afterwards;
Three, the bush mycorrhizal fungi preparation culture matrix being put into is the culture pond of common brick for cement concrete, bottom all around, then treated host's seed and arbuscular mycorrhizal fungi are imposed on bush mycorrhizal fungi preparation culture matrix surface jointly, spray concentration is 5 of 150nmol/L again, 7,4'-trihydroxyflavone solution, cladding thickness is the bush mycorrhizal fungi preparation culture matrix of 0.4cm more afterwards; Wherein, the consumption of Apigenin solution is 250mL/ square metre; The culture pond degree of depth is 0.25 meter;
Four, conventional field management, be cultured to the cauline leaf part of removing host plant the 7th week, continue to cultivate to the main root of removing again host plant in the bush mycorrhizal fungi preparation culture matrix the 8th week, adventive root and bush mycorrhizal fungi preparation culture matrix mixing, granulation, namely obtain bush mycorrhizal fungi preparation; Wherein, every cubic metre of matrix is sprayed with the formalin dilution that 500mL contains formaldehyde in the step 1; In the step 2 length of culture pond be 5 meters, wide be 2 meters; Every square metre on bush mycorrhizal fungi preparation culture matrix surface adds mould 28000 spores of earth's surface sacculus in the step 3.
Adopt the acid fuchsin colouring method of Phillip and Hayman to measure Mycorrhizal Infection Incidence to Sorghum roots weekly, when the 8th week, adopt wet screening to microbial inoculum in fungal spore detect, testing result is as shown in table 4.
Table 4
| ? | The 2nd week | The 4th week | The 6th week | The 8th week |
| Mycorrhizal Infection Incidence (%) | 52.7±1.2 | 96.4±1.3 | 98.5±0.9 | 99.9±0.1 |
| Microbial inoculum miospore content (individual/g) | ? | ? | ? | 135±5 |
By to the statistical analysis of many groups sampled data, sorghum seedling is when growing into for the 2nd week, and Mycorrhizal Infection Incidence has reached more than 50%, and Mycorrhizal Infection Incidence is up to more than 95% when the 4th week.Particularly sacculus mould spore content in earth's surface shows that up to 135 spore content is abundant in the 8th when week every gram culture matrix.
Test:
Soil picks up from the agricultural land soil that Heilongjiang University's vegetables sample plot was never used atrazine, and its pH value is 6.35, organic carbon 11.46g/kg, rapid available phosphorus 124.23mg/kg, alkali-hydrolyzable nitrogen 196.34mg/kg, available potassium 131.76mg/kg, full nitrogen 9.18g/kg, full phosphorus (P
2O
5) 6.25g/kg, full potassium 15.42g/kg.Soil is not found indigenous AM fungal spore behind wet screening.
Test is the 5L plastic basin with container, adds in every basin for examination soil 3.0kg, water in the soil according to the agricultural chemicals spray that the content of atrazine (available from Heilongjiang Academy of Agricultural Sciences) concentration 50mg/kg will prepare, and continuous stirring and evenly mixing.30 of the sorghum seeds that sprouted of sowing (assorted No. 1 of dragon, Heilongjiang Academy of Agricultural Sciences provides) in each basin respectively, and add respectively embodiment 1,2, each 40g of bush mycorrhizal fungi preparation of 3 and 4, and set up the blank group.5 repetitions are established in each processing, amount to 25 basins, every basin final singling 16 strains.Water with weight method during the plant growth, guarantee that water in basin is not excessive, prevent that pollutant from outflowing with water.All basin alms bowl randomized arrangement are in Heilongjiang University scientific research greenhouse, after planting 20,40,60,80d, after rounding the mensuration that the strain plant carries out plant height and fresh weight, utilize its lateral root to calculate the mycorrhizal fungi infection rate of host plant, then residual quantity, microbiologic population and the soil enzyme activities of atrazine in the corresponding time rhizosphere soil are measured.
Experimental result
After making root compressing tablet and microscopy, each processes lower host plant Mycorrhizal Infection Incidence to calculate different time according to the Trouvelot method.The result shows, can both form mycorrhizas homobium with Chinese sorghum along with growth time prolongs the arbuscular mycorrhizal fungi that adds embodiment 1,2,3 and 4 bush mycorrhizal fungi preparations, and the Chinese sorghum infection rate is all presented the trend that increases gradually.When plant strain growth arrived 80d, Mycorrhizal Infection Incidence was all above 90%.
The Chinese sorghum that adds embodiment 1,2,3 and 4 bush mycorrhizal fungi preparations has reached more than the 5.02g at the in season weight average of 80d, is more than 2.3 times of blank group; And plant height is 1.4 times of blank group for reaching about 70.4cm.
Add the 60th day and the 80th day behind embodiment 1,2,3 and 4 bush mycorrhizal fungi preparations, the degradation rate of the atrazine in the soil is respectively 76.4%, 75.8%, 72.8%, 74.5% and 91.6%, 90.4%, 89.6%, 90.7%, all exceeds 50.2% and 61.3% of blank group.
Add embodiment 1,2,3 and 4 bush mycorrhizal fungi preparations to the obvious raising of urease activity in the atrazine-contaminated soil.
Mensuration adds four kinds of endogenous hormones (IAA---growth hormone, GA---gibberellin, ZR---zeatin and ABA---abscisic acid) level of symbiosis Chinese sorghum behind embodiment 1,2,3 and 4 bush mycorrhizal fungi preparations.The result is as shown in table 5.
Table 5
Bush mycorrhizal fungi preparation can improve the IAA level of plant seedlings; Bush mycorrhizal fungi preparation can significantly improve GA and ZR content the host plant seedling; Bush mycorrhizal fungi preparation on the ABA of host plant without impact.Experimental result illustrative experiment presentation of results improves the host plant Endogenous Hormones.
Claims (5)
1. the method for bush mycorrhizal fungi preparation is produced in batch production, it is characterized in that the method for batch production production bush mycorrhizal fungi preparation is carried out according to the following steps:
One, soil, sand and vermiculite are mixed into matrix by the volume ratio of 5:2:3,50 times of the formalin dilute with waters that will contain again 40% formaldehyde evenly are sprayed onto on the matrix afterwards, then with Polypropylence Sheet sealing 1 round the clock, spread out afterwards, insolation is 4～6 days in the sun, namely obtains the bush mycorrhizal fungi preparation culture matrix;
Two, Chinese sorghum, clover or trefoil seed are washed by rubbing with the hands 2～3 times with 20～30 ℃ of warm water, be that 0.3% liquor potassic permanganate soaked 3～4 hours with concentration again, then eluriate clean with clear water, afterwards again with 20～30 ℃ Seed soaking 24 hours and eluriate 2～3 times, host's seed treatment is finished in vernalization;
Three, the bush mycorrhizal fungi preparation culture matrix being put into is the culture pond of common brick for cement concrete, bottom all around, then treated host's seed and arbuscular mycorrhizal fungi are imposed on bush mycorrhizal fungi preparation culture matrix surface jointly, spray concentration is 5 of 150nmol/L again, 7,4'-trihydroxyflavone solution, cladding thickness is the bush mycorrhizal fungi preparation culture matrix of 0.4～0.7cm more afterwards; Wherein, the consumption of Apigenin solution is 250mL/ square metre; The culture pond degree of depth is 0.25～0.35 meter;
Four, conventional field management, be cultured to the cauline leaf part of removing host plant the 7th week, continue to cultivate to the main root of removing again host plant in the bush mycorrhizal fungi preparation culture matrix the 8th week, adventive root and bush mycorrhizal fungi preparation culture matrix mixing, granulation, namely obtain bush mycorrhizal fungi preparation.
2. the method for bush mycorrhizal fungi preparation is produced in batch production according to claim 1, it is characterized in that every cubic metre of matrix is sprayed with the formalin dilution that 500mL contains formaldehyde in the step 1.
3. the method for bush mycorrhizal fungi preparation is produced in batch production according to claim 1, the length that it is characterized in that culture pond in the step 2 be 5 meters, wide be 2 meters.
4. according to claim 1, the 2 or 3 described batch production method of producing bush mycorrhizal fungi preparations, it is characterized in that the arbuscular mycorrhizal fungi in the step 3 is that Glomus intraradices, Scotland sacculus are mould, the earth's surface sacculus is mould or Glomus mosseae.
5. the method for bush mycorrhizal fungi preparation is produced in batch production according to claim 4, it is characterized in that in the step 3 that every square metre on bush mycorrhizal fungi preparation culture matrix surface adds 25000～30000 spores of arbuscular mycorrhizal fungi.
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