CN106434389A - Penicillium sp. LX-ZJ-2 and application thereof - Google Patents
Penicillium sp. LX-ZJ-2 and application thereof Download PDFInfo
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Abstract
The invention relates to penicillium sp. LX-ZJ-2 and application thereof, in particular to application of the penicillium sp. LX-ZJ-2 in activation of silicon and calcium elements in water-quenched slags. The strain can obviously promote growth of plants when being applied together with the water-quenched slags and can promote root development when being applied to rice, and root system dry weight is increased by 34.52%; rice tiller number is increased by 36.35%; total potassium of the plants is increased by 6.4%; total nitrogen of the plants is increased by 9.6%; total phosphorus of the plants is increased by 9.4%; total silicon of the plants is increased by 13.6%. The strain can increase utilization rate of effective elements in the water-quenched slags when being applied together with the water-quenched slags, and has great application prospect in agricultural resource utilization of industrial residues such as residues from ironmaking and steelmaking.
Description
【Technical field】
The invention belongs to biologic applications technical field.More particularly it relates to one plant of penicillium sp (Penicillium
Sp.) LX-ZJ-2, further relates to the purposes of penicillium sp (Penicillium sp.) LX-ZJ-2.
【Background technology】
China is the first big producing country of world steel, up to 700,000,000 tons of Chinese crude steel yield in 2012, accounts for world steel production
The 46.3% of total amount.The side-product for discharging in ironmaking, steelmaking process, accounts for the 15～20% of iron and steel output.But, China's steel
The utilization rate of ferrum industrial residue only about 10%, far below developed countries such as America and Europes.A large amount of waste residues for no processing arbitrarily are piled up, no
A large amount of soils are only occupied, and environmental pollution is brought, even more a kind of wasting of resources.National " 12 " demand for development steel-making, refining
The silicate waste byproduct cyclic utilization rate of ferrum output reaches more than 60%, and it is steel industry face that the resource of industrial residue is recycled
The urgent task for facing.These silicate waste byproduct contain the nutrient required for a large amount of plant growings, such as Ca (29%～
36%), Si (4%～12%), Fe (6%～27%), Mg (1.8%～10.2%) and the element such as a small amount of P, Mn, Cu, Zn,
While these silicate waste byproduct also have larger specific surface area and porosity, it is excellent calcium silicate fertilizer raw material and acid ground
Modifying agent.Slag calcium silicate fertilizer has administration in many countries such as Japanese, the European, U.S. and China, and research shows this industry
Waste residue is made calcium silicate fertilizer and can remarkably promote plant growth, improves resistivity of the plant to biological and abiotic stress, then
Improve crop yield.Therefore the waste residue that these steel-making, ironmaking are produced is made agricultural calcium silicate fertilizer and not only contributes to agricultural production, and
To steel industry, all there is important economic benefit and environmental benefit.
What in these scums, slag, content was higher is element silicon, and Gui world soil circle is listed in after nitrogen, phosphorus, potassium
The fourth-largest element fertilizer.It is mainly used in happiness silicon crop, the especially Oryza sativa L. such as Oryza sativa L., Semen Tritici aestivi, Semen Maydiss.Oryza sativa L. have silicic acid plant
The title of thing, on scarce tripoli earth, Applying silicon fertilizer can increase rice tillering number, improve and effectively harvest spike number, can also improve solid
Rate, promotes dry matter accumulation, and then increases yield, and play the role of to improve rice matter；In addition, the effect of siliceous fertilizer blast resisting is most aobvious
Write, increase Oryza sativa L. itself resistances against diseases are mainly shown as, even if Oryza sativa L. infects some diseases, it is also possible to make scab siliceousization, control
Scab development processed；Next to that BPH resistant rice variety fringe, banded sclerotial blight.
At present, the kind of siliceous fertilizer mainly has solubility in citric acid siliceous fertilizer, two big class of Water-soluble silicon fertilizer, and solubility in citric acid siliceous fertilizer refers to not dissolve in
Water and the siliceous fertilizer that can be absorbed by plants after being dissolved in acid, the siliceous fertilizer that field is applied is mainly the industrial residue of silicate, such as steel
Slag, scum, copper slag, aluminium metallurgy red mud, flyash-Si calcareous fertilisers and Phosphorus Slag and cement flue Dust and silicon-magnesium potash fertilizer etc., this fertilizer
General amount of application is larger, but absorbs excessively slowly, is not easy to be absorbed by plants mainly due to silicate therein, so how
Improve effective element utilization rate in siliceous fertilizer so as to which being faster easier to be absorbed and used by plants becomes in the urgent need to address asking
Topic.The agricultural resources of industrial residue are made using more efficient by biological method, to developing ecology civilization agricultural and iron and steel row
The environmentally friendly cycle development of industry has great importance.
【Content of the invention】
[technical problem to be solved]
It is an object of the invention to provide one plant of penicillium sp (Penicillium sp.) LX-ZJ-2.
It is a further object to provide the purposes of penicillium sp LX-ZJ-2.
[technical scheme]
The present invention is achieved through the following technical solutions.
The present invention relates to one plant of penicillium sp (Penicillium sp.) LX-ZJ-2, the bacterial strain exists on June 23rd, 2016
The preservation of China Committee for Culture Collection of Microorganisms's common micro-organisms center of Institute of Microorganism, Academia Sinica, its preserving number
For CGMCCNo.12628.
The invention further relates to purposes of the described penicillium sp LX-ZJ-2 bacterial strain in activation Water Quenching Slag silicon and calcium constituent.
A preferred embodiment of the invention is as follows the step of the activation method：
Prepared by A, penicillium sp LX-ZJ-2 spore powder
1. prepared by PDA culture medium:
Prepare containing 20g/l sucrose, 200g/l Rhizoma Solani tuber osi, 20g/l agar, nature pH aqueous solution, allow the aqueous solution in temperature
Sterilize at 121 DEG C of degree 30min, obtains PDA culture medium；
2. penicillium sp culture：
One ring penicillium sp LX-ZJ-2 of picking is inoculated in the PDA culture medium that 1. step is prepared, in constant incubator at 28 DEG C
Slant culture 3～4 days, obtains a kind of penicillium sp culture；
3. prepared by spore powder：
The penicillium sp culture that 2. one ring step of picking obtains turns and is seeded in the PDA culture medium that 1. step is prepared, in constant temperature
In incubator at 28 DEG C of temperature flat board culture 3～4 days, obtain penicillium sp LX-ZJ-2 spore powder, it again with the dry rice of 80～100 mesh
Grass meal absorption carrier mix homogeneously, allows penicillium sp LX-ZJ-2 spore powder to adsorb completely on dry rice straw powder；
4. spore powder count plate：
The spore powder for 3. step being obtained using conventional viable counts method carries out count plate, and the spore powder viable count is
2～3 × 109Individual/gram；
B, Water Quenching Slag pretreatment
Using disintegrating apparatus, Water Quenching Slag is crushed to 150～200 mesh；
C, activation processing
According to spore powder：Water Quenching Slag：The weight ratio of soil is 1：0.8～1.2：450～550, the work for first allowing step A obtain
Bacterium number is 2～3 × 109Individual/gram the water quenching ground-slag that obtains with step B of penicillium sp LX-ZJ-2 spore powder mix homogeneously, then with agricultural
Mixing with soil is uniform, so that penicillium sp LX-ZJ-2 viable count reaches 10 in the soil6Individual/gram；
Then add water toward in the mixture so as to moisten and place, sampling, detects water-soluble silicon and exchangeable calcium content.
According to another kind of preferred implementation of the present invention, described Water Quenching Slag is produced in ironmaking processes rich in silicon
The slag of hydrochlorate.
According to another kind of preferred implementation of the present invention, the agricultural soil be 2 mm sieve sieve under soil.
The present invention is described in more detail below.
The present invention relates to one plant of penicillium sp (Penicillium sp.) LX-ZJ-2, the bacterial strain exists on June 23rd, 2016
The preservation of China Committee for Culture Collection of Microorganisms's common micro-organisms center of Institute of Microorganism, Academia Sinica, its preserving number
For CGMCC No.12628.
The Penicillium of the penicillium sp strain LX-ZJ-2 category funguses of the present invention, is ploughed by Hebei province Qinhuangdao Funing area crops
Make isolated in the root system of plant surrounding soil in area.Concrete separation method is as follows：
Above-mentioned for 10g pedotheque and 90g sterilized water are added in triangular flask, concussion shakes up, and obtains 10-1Dilution soil
Suspension, then takes 10ml soil supension, adds 90ml sterilized water thereto, and concussion shakes up, and obtains 10-2Dilution soil hangs
Liquid, prepares 10 in the same way in succession-3、10-4、10-5Dilution soil supension.
By different dilution soil supensions be respectively coated containing bromothymol blue (0.25% concentration by weight, 1
～2ml/l) conventional PDA solid medium on, cultivate 4-6 days at 28 DEG C of temperature.
When thalline is grown in the culture medium containing bromothymol blue, if produce acid if flat board turn yellow, if produce alkali if
Flat board becomes blue, and picking flat board produces the bacterial strain of sour area maximum and isolated and purified.Wherein bacterial strain LX-ZJ-2 produces sour face on flat board
Product is maximum, so choosing the bacterium colony to carry out purification culture.
Colony characteristicses：Constant temperature culture is after 4～6 days at 28 DEG C of temperature for the bacterial strain, and bacterium colony front forms dirty-green spore, bacterium
The back side that falls is faint yellow, 30～50mm of diameter, conidium ellipse, and size is 4～6 μm of (length) × 3～4 μm (width), mitogenetic spore
Son is born in chain on sporophore, and conidiophore is colourless, have separation, and stigma is grown thickly in branch top.
Strain identification：The bacterial strain carries out 18S rDNA sequencing, according to known to those skilled in the art true
Bacterium DNA extraction method prepares sample total DNA.
Synthesis fungal DNA universal primer is as follows：
ITS1 5’-TCCGTAGGTGAACCTGCGG-3’
ITS4 5'-TCCTCCGCTTATTGATATGC-3’
PCR system (25 μ L)：
Response procedures：94℃4min；94 DEG C of 1min, 52 DEG C of 1min, 72 DEG C of 1min, 35 circulations；72℃10min.
Beijing three is sent to win the sequencing of Radix Polygalae company after agarose gel electrophoresiies detection is qualified.The DNA sequence for measuring is entered pedestrian
After work is corrected, using being compared in the Blast program in NCBI online, through comparing, the bacterial strain is 99% with the similarity of penicillium sp,
So concluding the bacterial strain for penicillium sp.
The invention further relates to the purposes of described penicillium sp LX-ZJ-2 bacterial strain silicon and calcium constituent in activation Water Quenching Slag.
Penicillium sp LX-ZJ-2 bacterial strain is as follows to silicon in Water Quenching Slag and the test of calcium activation：
The preparation of PDA culture medium:Prepare containing 20g/l sucrose, 200g/l Rhizoma Solani tuber osi, 20g/l agar, nature pH water-soluble
Liquid, allows the aqueous solution to sterilize at 121 DEG C of temperature 30min, obtains PDA culture medium；
The preparation of Ma Dingshi fluid medium：Prepare containing 10g/l glucose, 5g/l peptone, 1g/l potassium dihydrogen phosphate,
0.5g/l Magnesium sulfate heptahydrate, the aqueous solution of nature pH, allow the aqueous solution to sterilize at 112 DEG C of temperature 30min, obtain Ma Dingshi liquid
Body culture medium.
Penicillium sp culture：One ring penicillium sp LX-ZJ-2 of picking is inoculated in PDA culture medium, in temperature 28 in constant incubator
DEG C lower inclined plane culture 3～4 days, obtains a kind of penicillium sp culture；
The preparation of Water Quenching Slag：Water Quenching Slag is crushed to 150-200 mesh.
Two groups of experiment point is carried out, wherein：
First group be toward Ma Dingshi fluid medium in add Water Quenching Slag (0.5g/100ml)；
Second group be toward Ma Dingshi fluid medium in add Water Quenching Slag (0.5g/100ml) with a ring penicillium sp LX-ZJ-2；
Parallel carry out according to three per group；
Two groups of cultures being all put in constant-temperature table 48h is cultivated at 28 DEG C of temperature, is then filtered with filter paper, obtain
Supernatant using the water-soluble silicon content in conventional molybdenum blue colorimetric method detection fermentation liquid.
Testing result shows, first group of no water-soluble silicon content of penicillium sp LX-ZJ-2 is 816.83mg/kg；Second group has
The water-soluble silicon content of penicillium sp LX-ZJ-2 is 2369.35mg/kg；Second group of water-soluble silicon content is higher than first group by 190%.
This experiment fully demonstrates penicillium sp LX-ZJ-2 and can clearly be effectively facilitated Water Quenching Slag release water-soluble silicon.
According to the present invention, the step of penicillium sp LX-ZJ-2 activates silicon and calcium constituent method in Water Quenching Slag in soil environment such as
Under：
Prepared by A, penicillium sp LX-ZJ-2 spore powder
1. prepared by PDA culture medium:
Prepare containing 20g/l sucrose, 200g/l Rhizoma Solani tuber osi, 20g/l agar, nature pH aqueous solution, allow the aqueous solution in temperature
Sterilize at 121 DEG C of degree 30min, obtains PDA culture medium；
2. penicillium sp culture：
One ring penicillium sp LX-ZJ-2 of picking is inoculated in the PDA culture medium that 1. step is prepared, in temperature in constant incubator
28 DEG C of lower inclined plane cultures 3～4 days, obtain a kind of penicillium sp culture；
3. prepared by spore powder：
The penicillium sp culture that 2. one ring step of picking obtains turns and is seeded in the PDA culture medium that 1. step is prepared, in constant temperature
In incubator at 28 DEG C of temperature flat board culture 3～4 days, obtain penicillium sp LX-ZJ-2 spore powder, it again with the dry rice of 80～100 mesh
Grass meal absorption carrier mix homogeneously, allows penicillium sp LX-ZJ-2 spore powder to adsorb completely on dry rice straw powder；
4. spore powder count plate：
The spore powder for 3. step being obtained using conventional viable counts method carries out count plate, and the spore powder viable count is
2～3 × 109Individual/gram；
B, Water Quenching Slag pretreatment
Using disintegrating apparatus, Water Quenching Slag is crushed to 150～200 mesh；Described Water Quenching Slag is blast furnace heat in ironmaking processes
Molten slag water can be changed into the loose slag rich in silicate after rapidly cooling down.
C, activation processing
According to spore powder：Water Quenching Slag：The weight ratio of soil is 1：0.8～1.2：450～550, the work for first allowing step A obtain
Bacterium number is 2～3 × 109Individual/gram the water quenching ground-slag that obtains with step B of penicillium sp LX-ZJ-2 spore powder mix homogeneously, then with cross 2
Agricultural soil (lower part of screen is divided) mix homogeneously of mm sieve, so that penicillium sp LX-ZJ-2 viable count reaches 10 in the soil6Individual/
Gram；
Then add water toward in the mixture so as to moisten and place, sampling, detects water-soluble silicon and exchangeable calcium content.
Specifically, two groups of Water Quenching Slag soil water experiments are abreast carried out：
First group, the conventional soil of 500g is weighed, is stirred with 1g Water Quenching Slag, 100ml water is added, room temperature is placed, and is kept
Moistening；
Second group, the conventional soil of 500g is weighed, (spore powder viable count is 2 with 1g penicillium sp LX-ZJ-2 spore powder with 1g Water Quenching Slag
～3 × 109Individual/gram), stir, plus 100ml water, room temperature is placed, and keeps moistening；
Above-mentioned two groups of samples are placed 12 days, and soil sampling is dried in the shade, naturally using conventional molybdenum blue colorimetric method and soil exchangeable calcium
Magnesium detection method, detects the water-soluble silicon in these pedotheques and exchangeable calcium.Testing result is listed in the table below in 1.
Table 1：Water-soluble silicon and exchangeable calcium testing result in pedotheque
| First group | Second group | Improve % | |
| Exchangeable calcium, g/kg | 7.16 | 7.81 | 9% |
| Water-soluble silicon, g/kg | 232.44 | 276.56 | 19% |
The result of table 1 shows, compared with penicillium sp LX-ZJ-2 spore powder is not applied, applies penicillium sp LX-ZJ-2 spore powder
Exchangeable calcium improves 9%, and water-soluble silicon improves 19%, and this result absolutely proves that penicillium sp LX-ZJ-2 spore powder can be very bright
Aobvious promotion Water Quenching Slag release silicon and calcium, therefore have very important significance.
Penicillium sp LX-ZJ-2 and Water Quenching Slag are as follows in Rice Cropping application test：
I, test soil：Qinhuangdao Funing rice soil, it contain 59.7mg/kg alkali-hydrolyzable nitrogen, 18.6mg/kg available phosphoruss,
The effective potassium of 117.2mg/kg, 2.4% organic matter, the effective silicon of 110mg/kg, 30.6mg/kg soluble silicon, pH 5.8；Dry in the shade, choosing
Divided with the lower part of screen that is sieved by 2mm.
II, saturated soil water content are determined：
Take a certain amount of test soil to be put in Bush's funnel, constantly add water to and no longer drip substantially, overnight.Take test soil to put
Enter and weigh in aluminum box, with ethanol calcination to dry, be re-weighed.Water content is calculated according to soil weight before and after test, measures the Oryza sativa L.
Native saturation moisture content is by weight 37%.
III, test process：
CK：Base manure, per kg, soil is 0.15g N, 0.1g P2O5With 0.15g K2O.
Process 1：Base manure+penicillium sp LX-ZJ-2 spore powder (106Individual/kg soil)
Process 2：Base manure+Water Quenching Slag (2.3g/kg soil)
Process 3：Base manure+penicillium sp LX-ZJ-2 spore powder (106Individual/kg soil)+Water Quenching Slag (2.3g/kg soil)
Penicillium sp LX-ZJ-2 spore powder is the penicillium sp LX-ZJ-2 spore powder of carrier adsorption.
IV, detection project：
Full nitrogen, phosphorus, potassium, silicone content are detected using the conventional method of the art.
Seedling stage：Total nitrogen, phosphorus, potassium, silicone content；
Tillering stage：Total nitrogen, phosphorus, potassium, silicone content；
Period of maturation：Total nitrogen, phosphorus, potassium, silicone content；Tiller number, root weight, yield.
Result of the test is shown in Table 2-4.
Table 2：Rice root, fringe, tiller number measured value
Table 3：The full phosphorus of rice plant, total silicon measured value
Table 4：The full phosphorus of rice plant, total silicon measured value
By table 2-4 it can be clearly seen that when penicillium sp LX-ZJ-2 is with Water Quenching Slag while when applying, can be obviously promoted plant
Growth, improve utilization rate than Water Quenching Slag is administered alone, promote root development, improve weight of root system 34.52%；Improve Oryza sativa L.
Tiller number 36.35%；The full potassium of plant improves 6.4%；Total nitrogen improves 9.6%；The full phosphorus of plant improves 9.4%；Plant total silicon
Improve 13.6%.Therefore, by penicillium sp LX-ZJ-2 with Water Quenching Slag while application effect is highly significant, and people are not pre-
Expect, its popularization and application tool is of great significance.
[beneficial effect]
The invention has the beneficial effects as follows：The invention provides one plant of penicillium sp strain LX-ZJ-2, passes through it is demonstrated experimentally that the bacterial strain
The silicon in Water Quenching Slag and calcium can be activated, 48h under liquid environment, in Water Quenching Slag, water-soluble silicon burst size is improved than control
190%；The release rate for improving silicon and calcium in Water Quenching Slag can also be activated in soil environment, by Water Quenching Slag and penicillium sp strain LX-ZJ-
The release of Water Quenching Slag silicon and calcium in soil during 2 spore powders are manured into soil simultaneously, is investigated after 12 days, as a result adds the water of spore powder
Slag ratio of quenching improves 8.6～10.3% without the process of spore powder, exchangeable calcium release, and the release of water-soluble silicon improves
17.7～20.9%；
The bacterial strain applies the growth that can be obviously promoted plant simultaneously with Water Quenching Slag, improves Water Quenching Slag utilization rate, in Oryza sativa L.
After upper administration, root development can be promoted, improve weight of root system 34.52%；Improve rice tillering number 36.35%；The full potassium of plant
Improve 6.4%；Total nitrogen improves 9.6%；The full phosphorus of plant improves 9.4%；Plant total silicon improves 13.6%.
Therefore, the bacterial strain with Water Quenching Slag and meanwhile apply when can improve effective element utilization rate in Water Quenching Slag, ironmaking,
The industrial residue agricultural resources utilization orientation such as steel-making have extremely well application prospect.
Penicillium sp (Penicillium sp.) LX-ZJ-2, the bacterial strain is on June 23rd, 2016 in the micro- life of the Chinese Academy of Sciences
The preservation of thing institute China Committee for Culture Collection of Microorganisms's common micro-organisms center, its preserving number is
CGMCCNo.12628.
【Specific embodiment】
The present invention be will be better understood that by following embodiments.
Embodiment 1：Silicon and calcium constituent activation processing in Water Quenching Slag
The implementation steps of the embodiment are as follows：
Prepared by A, penicillium sp LX-ZJ-2 spore powder
1. prepared by PDA culture medium:
Prepare containing 20g/l sucrose, 200g/l Rhizoma Solani tuber osi, 20g/l agar, nature pH aqueous solution, allow the aqueous solution in temperature
Sterilize at 121 DEG C of degree 30min, obtains PDA culture medium；
2. penicillium sp culture：
One ring penicillium sp LX-ZJ-2 of picking is inoculated in the PDA culture medium that 1. step is prepared, in temperature in constant incubator
28 DEG C of lower inclined plane cultures 3 days, obtain a kind of penicillium sp culture；
3. prepared by spore powder：
The penicillium sp culture that 2. one ring step of picking obtains turns and is seeded in the PDA culture medium that 1. step is prepared, in constant temperature
In incubator at 28 DEG C of temperature flat board culture 4 days, obtain penicillium sp LX-ZJ-2 spore powder, it is adsorbed with 80 mesh dry rice straw powder again
Carrier mix homogeneously, allows penicillium sp LX-ZJ-2 spore powder to adsorb completely on dry rice straw powder；
4. spore powder count plate：
The PDA plate culture spore powder for 3. step being obtained using conventional viable counts method carries out count plate, should
Spore powder viable count is 2 × 109Individual/gram；
B, Water Quenching Slag pretreatment
Using disintegrating apparatus, Water Quenching Slag is crushed to 180 mesh；
C, activation processing
According to spore powder：Water Quenching Slag：The weight ratio of soil is 1：0.8：550, the viable count for first allowing step A obtain be 2 ×
109Individual/gram the water quenching ground-slag that obtains with step B of penicillium sp LX-ZJ-2 spore powder mix homogeneously, then with cross the Qin emperor of 2 mm sieve
Island Funing rice soil mix homogeneously, so that penicillium sp LX-ZJ-2 viable count reaches 10 in the soil6Individual/gram；
Two group Water Quenching Slag soil water experiments are abreast carried out below：
First group, the conventional soil of 550g is weighed, is stirred with 0.8g Water Quenching Slag, 100ml water is added, room temperature is placed, and is protected
Hold moistening；
Second group, the conventional soil of 550g is weighed, with 0.8g Water Quenching Slag and 1g penicillium sp LX-ZJ-2 spore powder (spore powder viable count
For 2 × 109Individual/gram), stir, plus 100ml water, room temperature is placed, and keeps moistening；
Above-mentioned two groups of samples are placed 12 days, and soil sampling is dried in the shade, naturally using conventional molybdenum blue colorimetric method and soil exchangeable calcium
Magnesium detection method, detects the water-soluble silicon in these pedotheques and exchangeable calcium.The results are shown in Table 5 for which.
Table 5：Water-soluble silicon and exchangeable calcium testing result in pedotheque
| First group | Second group | Improve % | |
| Exchangeable calcium, g/kg | 7.18 | 7.92 | 10.3% |
| Water-soluble silicon, g/kg | 229.44 | 277.61 | 20.9% |
Embodiment 2：Silicon and calcium constituent activation processing in Water Quenching Slag
The implementation steps of the embodiment are as follows：
Prepared by A, penicillium sp LX-ZJ-2 spore powder
1. prepared by PDA culture medium:
Prepare containing 20g/l sucrose, 200g/l Rhizoma Solani tuber osi, 20g/l agar, nature pH aqueous solution, allow the aqueous solution in temperature
Sterilize at 121 DEG C of degree 30min, obtains PDA culture medium；
2. penicillium sp culture：
One ring penicillium sp LX-ZJ-2 of picking is inoculated in the PDA culture medium that 1. step is prepared, in temperature in constant incubator
28 DEG C of lower inclined plane cultures 4 days, obtain a kind of penicillium sp culture；
3. prepared by spore powder：
The penicillium sp culture that 2. one ring step of picking obtains turns and is seeded in the PDA culture medium that 1. step is prepared, in constant temperature
In incubator at 28 DEG C of temperature flat board culture 3 days, obtain penicillium sp LX-ZJ-2 spore powder, it is adsorbed with 100 mesh dry rice straw powder again
Carrier mix homogeneously, allows penicillium sp LX-ZJ-2 spore powder to adsorb completely on dry rice straw powder；
4. spore powder count plate：
The PDA plate culture spore powder for 3. step being obtained using conventional viable counts method carries out count plate, should
Spore powder viable count is 3 × 109Individual/gram；
B, Water Quenching Slag pretreatment
Using disintegrating apparatus, Water Quenching Slag is crushed to 150 mesh；
C, activation processing
According to spore powder：Water Quenching Slag：The weight ratio of soil is 1：1.0：450, the viable count for first allowing step A obtain be 3 ×
109Individual/gram the water quenching ground-slag that obtains with step B of penicillium sp LX-ZJ-2 spore powder mix homogeneously, then with cross the Qin emperor of 2 mm sieve
Island Funing rice soil mix homogeneously, so that penicillium sp LX-ZJ-2 viable count reaches 10 in the soil6Individual/gram；
Two group Water Quenching Slag soil water experiments are abreast carried out below：
First group, the conventional soil of 450g is weighed, is stirred with 1g Water Quenching Slag, 100ml water is added, room temperature is placed, and is kept
Moistening；
Second group, the conventional soil of 450g is weighed, (spore powder viable count is 3 with 1g penicillium sp LX-ZJ-2 spore powder with 1g Water Quenching Slag
×109Individual/gram), stir, plus 100ml water, room temperature is placed, and keeps moistening；
Above-mentioned two groups of samples are placed 12 days, and soil sampling is dried in the shade, naturally using conventional molybdenum blue colorimetric method and soil exchangeable calcium
Magnesium detection method, detects the water-soluble silicon in these pedotheques and exchangeable calcium.The results are shown in Table 6 for which.
Table 6：Water-soluble silicon and exchangeable calcium testing result in pedotheque
| First group | Second group | Improve % | |
| Exchangeable calcium, g/kg | 6.99 | 7.59 | 8.6% |
| Water-soluble silicon, g/kg | 235.71 | 283.02 | 20.5% |
Embodiment 3：Silicon and calcium constituent activation processing in Water Quenching Slag
The implementation steps of the embodiment are as follows：
Prepared by A, penicillium sp LX-ZJ-2 spore powder
1. prepared by PDA culture medium:
Prepare containing 20g/l sucrose, 200g/l Rhizoma Solani tuber osi, 20g/l agar, nature pH aqueous solution, allow the aqueous solution in temperature
Sterilize at 121 DEG C of degree 30min, obtains PDA culture medium；
2. penicillium sp culture：
One ring penicillium sp LX-ZJ-2 of picking is inoculated in the PDA culture medium that 1. step is prepared, in temperature in constant incubator
28 DEG C of lower inclined plane cultures 3 days, obtain a kind of penicillium sp culture；
3. prepared by spore powder：
The penicillium sp culture that 2. one ring step of picking obtains turns and is seeded in the PDA culture medium that 1. step is prepared, in constant temperature
In incubator at 28 DEG C of temperature flat board culture 4 days, obtain penicillium sp LX-ZJ-2 spore powder, it is adsorbed with 90 mesh dry rice straw powder again
Carrier mix homogeneously, allows penicillium sp LX-ZJ-2 spore powder to adsorb completely on dry rice straw powder；
4. spore powder count plate：
The PDA plate culture spore powder for 3. step being obtained using conventional viable counts method carries out count plate, should
Spore powder viable count is 2.6 × 109Individual/gram；
B, Water Quenching Slag pretreatment
Using disintegrating apparatus, Water Quenching Slag is crushed to 200 mesh；
C, activation processing
According to spore powder：Water Quenching Slag：The weight ratio of soil is 1：1.2：550, the viable count for first allowing step A obtain is 2.6
×109Individual/gram the water quenching ground-slag that obtains with step B of penicillium sp LX-ZJ-2 spore powder mix homogeneously, then with cross the Qin of 2 mm sieve
Huang Dao Funing rice soil mix homogeneously, so that penicillium sp LX-ZJ-2 viable count reaches 10 in the soil6Individual/gram；
Two group Water Quenching Slag soil water experiments are abreast carried out below：
First group, the conventional soil of 500g is weighed, is stirred with 1.2g Water Quenching Slag, 100ml water is added, room temperature is placed, and is protected
Hold moistening；
Second group, the conventional soil of 500g is weighed, with 1.2g Water Quenching Slag and 1g penicillium sp LX-ZJ-2 spore powder (spore powder viable count
For 2.6 × 109Individual/gram), stir, plus 100ml water, room temperature is placed, and keeps moistening；
Above-mentioned two groups of samples are placed 12 days, and soil sampling is dried in the shade, naturally using conventional molybdenum blue colorimetric method and soil exchangeable calcium
Magnesium detection method, detects the water-soluble silicon in these pedotheques and exchangeable calcium.The results are shown in Table 7 for which.
Table 7：Water-soluble silicon and exchangeable calcium testing result in pedotheque
| First group | Second group | Improve % | |
| Exchangeable calcium, g/kg | 6.88 | 7.49 | 8.9% |
| Water-soluble silicon, g/kg | 232.61 | 273.67 | 17.7% |
Claims (5)
1. one plant of penicillium sp (Penicillium sp.) LX-ZJ-2, the bacterial strain is micro- in the Chinese Academy of Sciences on June 23rd, 2016
The preservation of China Committee for Culture Collection of Microorganisms's common micro-organisms center of biological study institute, its preserving number is
CGMCCNo.12628.
2. penicillium sp LX-ZJ-2 bacterial strain according to claim 1 is activating Water Quenching Slag silicon and the purposes in calcium constituent.
3. purposes according to claim 2, it is characterised in that as follows the step of the activation method：
Prepared by A, penicillium sp LX-ZJ-2 spore powder
1. prepared by PDA culture medium:
Prepare containing 20g/l sucrose, 200g/l Rhizoma Solani tuber osi, 20g/l agar, nature pH aqueous solution, allow the aqueous solution in temperature 121
Sterilize at DEG C 30min, obtains PDA culture medium；
2. penicillium sp culture：
One ring penicillium sp LX-ZJ-2 of picking is inoculated in the PDA culture medium that 1. step is prepared, in 28 DEG C of temperature in constant incubator
Lower inclined plane culture 3～4 days, obtains a kind of penicillium sp culture；
3. prepared by spore powder：
The penicillium sp culture that 2. one ring step of picking obtains turns and is seeded in the PDA culture medium that 1. step is prepared, in constant temperature culture
In case at 28 DEG C of temperature flat board culture 3～4 days, obtain penicillium sp LX-ZJ-2 spore powder, it again with 80～100 mesh dry rice straw powder
Absorption carrier mix homogeneously, allows penicillium sp LX-ZJ-2 spore powder to adsorb completely on dry rice straw powder；
4. spore powder count plate：
The PDA plate culture spore powder for 3. step being obtained using conventional viable counts method carries out count plate, the spore
Powder viable count is 2～3 × 109Individual/gram；
B, Water Quenching Slag pretreatment
Using disintegrating apparatus, Water Quenching Slag is crushed to 150～200 mesh；
C, activation processing
According to spore powder：Water Quenching Slag：The weight ratio of soil is 1：0.8～1.2：450～550, the viable count for first allowing step A obtain
For 2～3 × 109Individual/gram the water quenching ground-slag that obtains with step B of penicillium sp LX-ZJ-2 spore powder mix homogeneously, then with agricultural soil
Mix homogeneously, so that penicillium sp LX-ZJ-2 viable count reaches 10 in the soil6Individual/gram；
Then add water toward in the mixture so as to moisten and place, sampling, detects the water-soluble silicon in its mixture and commutativity
Calcium content.
4. purposes according to claim 2, it is characterised in that described Water Quenching Slag is being rich in of producing in ironmaking processes
The slag of silicate.
5. purposes according to claim 2, it is characterised in that the agricultural soil be 2 mm sieve sieve under soil.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN114437941A (en) * | 2022-01-28 | 2022-05-06 | 领先生物农业股份有限公司 | Saline-alkali-resistant penicillium oxalicum agent, and preparation method and application thereof |
| CN116676201A (en) * | 2023-08-01 | 2023-09-01 | 河北省科学院生物研究所 | Penicillium poland CFP-2 and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102391021A (en) * | 2011-08-15 | 2012-03-28 | 广州钢铁企业集团有限公司 | Method for producing silicon-calcium-magnesium fertilizer by using ironmaking blast furnace water-quenching slag as raw materials |
| CN105838627A (en) * | 2016-05-10 | 2016-08-10 | 湖北省烟草公司恩施州公司 | Penicillium xfsf-8 strain for preventing and treating tobacco bacterial wilt and application |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102391021A (en) * | 2011-08-15 | 2012-03-28 | 广州钢铁企业集团有限公司 | Method for producing silicon-calcium-magnesium fertilizer by using ironmaking blast furnace water-quenching slag as raw materials |
| CN105838627A (en) * | 2016-05-10 | 2016-08-10 | 湖北省烟草公司恩施州公司 | Penicillium xfsf-8 strain for preventing and treating tobacco bacterial wilt and application |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114437941A (en) * | 2022-01-28 | 2022-05-06 | 领先生物农业股份有限公司 | Saline-alkali-resistant penicillium oxalicum agent, and preparation method and application thereof |
| CN114437941B (en) * | 2022-01-28 | 2024-02-02 | 领先生物农业股份有限公司 | Saline-alkali tolerant penicillium oxalicum agent and preparation method and application thereof |
| CN116676201A (en) * | 2023-08-01 | 2023-09-01 | 河北省科学院生物研究所 | Penicillium poland CFP-2 and application thereof |
| CN116676201B (en) * | 2023-08-01 | 2023-11-14 | 河北省科学院生物研究所 | Penicillium poland CFP-2 and application thereof |
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