CN104086237A - Special biological phosphate fertilizer for crops and preparation method thereof - Google Patents
Special biological phosphate fertilizer for crops and preparation method thereof Download PDFInfo
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- CN104086237A CN104086237A CN201410304205.9A CN201410304205A CN104086237A CN 104086237 A CN104086237 A CN 104086237A CN 201410304205 A CN201410304205 A CN 201410304205A CN 104086237 A CN104086237 A CN 104086237A
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Abstract
The invention relates to a special biological phosphate fertilizer for crops which is a phosphorus-solubilizing fungus ground phosphorite fermentation liquid. The phosphorus-solubilizing fungus Aspergillus niger XXR-B1, Penicillium sp.XXR-G or Fungal sp.XXR-E is respectively prepared into the special biological phosphate fertilizer for corn, soybeans or mung beans. The preparation method comprises the following steps: carrying out phosphorus-solubilizing fungus culture on the soil sample, separating and purifying to obtain the phosphorus-solubilizing fungus, putting the phosphorus-solubilizing fungus in a ground phosphorite fermentation liquid, and fermenting and culturing at 28-30 DEG C for 15-18 days to obtain the phosphorus-solubilizing fungus ground phosphorite fermentation liquid which is the special biological phosphate fertilizer. The biological phosphate fertilizer avoids the problems of environmental pollution and food pollution caused by the mineral phosphate fertilizer, overcomes the defect of lack of special biological phosphate fertilizers at present, sufficiently utilizes the phosphorus-solubilizing microbe resources in the natural world, and provides better technical insurance for yield increase of crops.
Description
Technical field
The present invention relates to biologic phosphorus fertilizer bacterium, especially a kind of tailored version biologic phosphorus fertilizer preparation method.
Background technology
Low main of phosphate fertilizer utilization efficiency fixedly caused in soil by phosphate fertilizer.In order to meet the demand of agriculture production to phosphate fertilizer, people have strengthened the dynamics of phosphorate rock mining, but ecotope have been caused to irreparable damage:
phosphorate rock mining is to environment havoc;
the waste water of discharging in phophatic fertilizer production process;
mineral phosphate fertilizer contains some heavy metal (as cadmium etc.)/radioactive substance and causes food pollution.The life-time service of chemical fertilizer, causes chemical substance residual in a large number in soil, has destroyed organic composition, the eubiosis of soil, occurs salting of soil phenomenon, the grain yield drops.Advocate now the ecological agriculture, Development of Green Food, require people to use bio-feritlizer and replace chemical fertilizer, therefore, biologic phosphorus fertilizer arises at the historic moment.
Biologic phosphorus fertilizer is the bacteria preparation that contains obtaining by solid or liquid submerged fermentation, flora is mainly molten phosphorus microorganism, this microorganism not only can dissolution of minerals phosphorus, also can dissolve the water-insoluble phosphorus in soil, make it to be converted into can utilization absorbed by crops biologically effective phosphorus.Biologic phosphorus fertilizer has following advantage:
the equilibrium of fertilizer speed is moderate, and fertilizer efficiency is lasting, is difficult for by soil fixing and loss, and P use efficiency is higher than mineral phosphate fertilizer;
long-term application, is conducive to promote soil body humify, improves soil water-reataining fertilizer-keeping ability;
effectively prevent root-rot, rotten is waited root diseases, reduces the usage quantity of agricultural chemicals, improves crop quality.
Yet biologic phosphorus fertilizer in the market lacks crop tailored version, had a strong impact on the fertilizer efficiency of biologic phosphorus fertilizer, be unfavorable for crop yield, even the molten phosphorus microorganism in some biologic phosphorus fertilizers also may suppress plant growth, causes on the contrary crop yield to decline.Between microorganism and crop, exist significant interaction, what have can promote plant growth, and what have can suppress plant growth; Meanwhile, root exudates kind and the quantity variance of Different Crop are remarkable, surely grow and grow, and then affect molten phosphorus ability thereby affect phosphorus-solubilizing bacteria in soil.So screening has good molten phosphorus ability, field planting energy for growth, promotes the molten phosphorus microorganism of plant growth, produces crop tailored version biologic phosphorus fertilizer, is the development trend of biologic phosphorus fertilizer, is also to avoid one of Main Means that current biologic phosphorus fertilizer fertilizer efficiency is not high.A kind of tailored version biologic phosphorus fertilizer preparation method provided by the invention, adopt potted plant mode to test crop tailored version biologic phosphorus fertilizer effect of increasing production, can obtain convenient, fast, exactly the crop tailored version biologic phosphorus fertilizer that promotes specific plant growth and improve output, the unfavorable factors such as environmental pollution that mineral phosphate fertilizer causes, food contamination have been avoided, made up the weak point that current biologic phosphorus fertilizer lacks tailored version simultaneously, natural molten phosphorus Microbial resources have fully been excavated, for increasing crop yield provides better technical guarantee.
Summary of the invention
Technical problem to be solved by this invention is to adopt potted plant mode to test tailored version biologic phosphorus fertilizer effect of increasing production, and a kind of crop tailored version biologic phosphorus fertilizer and preparation method thereof is provided.
Tailored version biologic phosphorus fertilizer of the present invention is molten phosphorus fungi ground phosphate rock fermented liquid, and molten phosphorus fungi is
aspergillus nigerxXR-B1, during the GenBank number of logging in KF572126, makes corn tailored version biologic phosphorus fertilizer.
Another kind of tailored version biologic phosphorus fertilizer provided by the invention, this biologic phosphorus fertilizer is molten phosphorus fungi ground phosphate rock fermented liquid, molten phosphorus fungi is
penicilliumsp. XXR-G, during the GenBank number of logging in KF572132, makes soybean tailored version biologic phosphorus fertilizer.
The present invention also provides a kind of tailored version biologic phosphorus fertilizer, and this biologic phosphorus fertilizer is molten phosphorus fungi ground phosphate rock fermented liquid, and molten phosphorus fungi is Fungal sp. XXR-E, during the GenBank number of logging in KF572130, makes mung bean tailored version biologic phosphorus fertilizer.
A kind of tailored version biologic phosphorus fertilizer preparation method provided by the present invention, the method comprises the following steps:
A preparation method for tailored version biologic phosphorus fertilizer, is characterized in that, comprises the following steps:
1. gather soil sample, and this soil sample is inoculated in to phosphorus-solubilizing bacteria screening culture medium after gradient dilution
In, at 25 ℃-30 ℃, cultivate 5-10 days, make the fungal colony edge in soil sample occur molten phosphorus circle, the fungal colony that produces molten phosphorus circle is forwarded in new phosphorus-solubilizing bacteria screening culture medium, at 25 ℃-30 ℃, cultivate 3-5 days, carry out obtaining molten phosphorus fungi after separation and purification.Described soil sample is 10 through gradient dilution
-1, 10
-2, 10
-3.Described phosphorus-solubilizing bacteria screening culture medium is that glucose, calcium phosphate, 6 aqueous magnesium chlorides, 7 water magnesium sulfates, Repone K, ammonium sulfate, agar be take to mass ratio as 15:5:5:0.25:0.2:0.1:15 mixing.
2. the molten phosphorus fungi after separation and purification is placed in to ground phosphate rock fermented liquid, 28 ℃ of-30 ℃ of bottom fermentations trainings
Support 15-18 days, obtain molten phosphorus fungi ground phosphate rock fermented liquid, this molten phosphorus fungi ground phosphate rock fermented liquid is tailored version biologic phosphorus fertilizer.Described ground phosphate rock fermented liquid is by weight mixing for 10:10:5:0.25:0.2:0.1 by glucose, ground phosphate rock, 6 aqueous magnesium chlorides, 7 water magnesium sulfates, Repone K, ammonium sulfate.
The screening of tailored version biologic phosphorus fertilizer has particular application as:
From planting site, fetch earth, be screened to 6mm, by the soil sieving and the agricultural crop straw that is crushed to 5-10mm
Mix, spray water to the mixture water content of soil stalk be 40-50%, in soil stalk mixture, add step 2 again) in molten phosphorus fungi ground phosphate rock fermented liquid, and the soil that this is mixed is placed in the cultivation bucket that sets in advance cultivation matrix, as cultivating soil; Described cultivating soil and the mass ratio of stalk are 8:1, and the amount of adding molten phosphorus fungi ground phosphate rock fermented liquid is 1 * 10
5-9 * 10
5cFU/ gram of soil; Cultivation matrix packs in cultivation bucket after 20-250kg soil being crossed to the aperture of 2cm, stocky, reaches 4/5 height of cultivation bucket.
Culture underglass soil on the cultivation matrix soil of cultivation bucket (bucket internal upper part 1/5 height), spray water to soil moisture content be 60-75%, and crop seed kind is screened in the cultivating soil layer at bucket internal upper part 1/5 place to suitable tailored version biologic phosphorus fertilizer and produces bacterial classification.
A kind of tailored version biologic phosphorus fertilizer preparation method provided by the invention, adopt potted plant mode to test tailored version biologic phosphorus fertilizer effect of increasing production, can obtain convenient, fast, exactly the crop tailored version biologic phosphorus fertilizer that promotes plant growth and improve output, the unfavorable factors such as environmental pollution that mineral phosphate fertilizer causes, food contamination have been avoided, made up the weak point that current biologic phosphorus fertilizer lacks tailored version simultaneously, natural molten phosphorus Microbial resources have fully been excavated, for increasing crop yield provides better technical guarantee.
Embodiment
Embodiment mono-
A tailored version biologic phosphorus fertilizer preparation method, the method comprises the following steps:
1) from different crop-plantings gather 57 of soil samples.
2) cultivate: soil sample is done to 10
-1, 10
-2, 10
-3after serial dilution, be inoculated in every liter of phosphorus-solubilizing bacteria screening culture medium NBRIP(NBRIP and contain: 10 g glucose, 5 g Ca
3(PO
4)
2, 5 g MgCl
26H2O, 0.25 g MgSO
47H
2o, 0.2 g KCl, 0.1 g (NH
4)
2sO
4, 15 g agar) in, cultivate 10 days for 28 ℃.
3) observe molten phosphorus circle: molten phosphorus circle appears in 10 fungal colonization edges.
4) separation and purification: the bacterium colony that step 3) is produced to molten phosphorus circle is forwarded to new phosphorus-solubilizing bacteria screening culture medium NBRIP above, carries out separation and purification, obtains the molten phosphorus fungi of 8 strain.
5) will when NBRIP liquid fermenting, (be fungi by the molten phosphorus fungi of 4 strain of activity high [fermented liquid phosphorus content >25 μ g/ml], be numbered Z01, Z02, Z03 and Z04), with ground phosphate rock fermented liquid [ground phosphate rock fermented liquid, every liter contains: 10 g glucose, 10 g ground phosphate rock are (containing P
2o
526%), 5 g MgCl
26H2O, 0.25 g MgSO
47H
2o, 0.2 g KCl, 0.1 g (NH
4)
2sO
4)], enlarged culturing, cultivates 16 days for 28 ℃; Control group is not for adding molten phosphorus fungi ground phosphate rock fermented liquid.
6) selecting cultivation bucket diameter is 40cm, dark 60cm, and bottom of the barrel borehole diameter 3cm is beneficial under excessive moisture and oozes.
7) cultivation bucket is put into the heatable adobe sleeping platform digging in advance, plane 5cm above ground level bucket on, then with the native hole tamping barrel with surrounding soil.
8) soil heatable adobe sleeping platform being dug out is removed macrobead with 2cm aperture particle size stainless (steel) wire, and after repeatedly mixing, as cultivation matrix, every barreled carries 150kg, steady and sure 4/5 height to cultivation bucket.
9) from planting site fetch earth (water content is 40-50%), with sieve aperture 6mm stainless (steel) wire, remove macrobead, the soil sieving is mixed with the corn cob of pulverizing, by soil: corn cob=8:1 (w/w), fully mix (corn cob water content is 40-50%), mixed soil is mixed with molten phosphorus fungi ground phosphate rock fermented liquid to (bacteria containing amount is 7.0 * 10
5cfu/g is native), or contrast ground phosphate rock nutrient solution mixes.The soil that contains phosphorus-solubilizing bacteria or contrast culture liquid is contained in to cultivation matrix top in cultivation bucket, reach cultivation bucket top edges (cultivation bucket 1/5 position, top), during plantation, making soil moisture content is 60-75%, records planting soil available phosphorus content 14.52 mg/kg.
10) corn seed (middle agricultural university 451) is planted in the soil of bucket internal upper part 1/5, after emerging, each barrel stays the seedling of 1 strain stalwartness.During results, control group soil available phosphorus content 5.034 ± 0.214 mg/kg (n=10), with other phosphorus-solubilizing bacteria group comparison, the highest (9.942 ± 0.421 mg/kg of Z02 barrel plant soil available phosphorus content, n=10), in soil, the quantity of Z02 is also the highest [(4.41 ± 0.26) * 10
4cfu/g soil, n=10]; Corn individual plant is dried artifact output (comprising root weight, cauline leaf weight, fruit ear weight) and fringe grain focuses on Z02 treatment group the highest (biological yield 342.86 ± 6.44 g/ strains, n=10; Fringe grain weighs 134.39 ± 2.07g/ strain, n=10), and compare with other group significant difference (
p<0.05).Biological yield and fringe grain weight-average value are respectively than control group (biological yield 290.65 ± 5.81g/ strain, n=10; Fringe grain weighs 107.27 ± 2.41g/ strain, n=10) improves 17.96% and 25.28%.To the Molecular Identification of molten phosphorus fungi Z02 find with aspergillus niger (
aspergillus niger) similarity reaches 99%, called after
aspergillus nigerxXR-B1, the GenBank number of logging in KF572126, corn tailored version biologic phosphorus fertilizer prepared by molten phosphorus fungi Z02, has obvious effect of increasing production.
Embodiment bis-
1) impact on soybean yields by the molten phosphorus fungi of 4 strain (being numbered Z01, Z02, Z03 and the Z04) test in embodiment mono-.
2) with ground phosphate rock fermented liquid, [ground phosphate rock fermented liquid, every liter contains: 10 g glucose, 10 g ground phosphate rock are (containing P
2o
526%), 5 g MgCl
26H2O, 0.25 g MgSO
47H
2o, 0.2 g KCl, 0.1 g (NH
4)
2sO
4)], enlarged culturing, cultivates 16 days for 28 ℃; Control group is not for adding molten phosphorus fungi ground phosphate rock fermented liquid.
3) selecting cultivation bucket diameter is 25cm, dark 40cm, and bottom of the barrel borehole diameter 2cm is beneficial under excessive moisture and oozes.
4) cultivation bucket is put into the heatable adobe sleeping platform digging in advance, plane 5cm above ground level bucket on, then with the native hole tamping barrel with surrounding soil.
5) soil heatable adobe sleeping platform being dug out is removed macrobead with 2cm aperture particle size stainless (steel) wire, after repeatedly mixing, as cultivation matrix, 40kg soil barrelling having peace of mind is extremely cultivated to 4/5 height of bucket.
6) from planting site fetch earth (water content is 40-50%), with sieve aperture 6mm stainless (steel) wire, remove macrobead, the soil sieving is mixed with the soybean cane of pulverizing, by soil: soybean cane=8:1 (w/w), fully mix (soybean cane water content is 40-50%), mixed soil is mixed with molten phosphorus fungi ground phosphate rock fermented liquid to (bacteria containing amount is 6.0 * 10
5cfu/g is native), or contrast ground phosphate rock nutrient solution mixes.The soil that contains phosphorus-solubilizing bacteria or contrast culture liquid is contained in to cultivation matrix top in cultivation bucket, reach cultivation bucket top edges (cultivation bucket 1/5 position, top), during plantation, making soil moisture content is 60-75%, records planting soil available phosphorus content 15.67 mg/kg.
7) soybean seeds (Nan Nong 38) is planted in the soil of bucket internal upper part 1/5, after emerging, each barrel stays the seedling of 2 strain stalwartnesses.During results, control group soil available phosphorus content 4.90 ± 0.13mg/kg (n=10), with other phosphorus-solubilizing bacteria group comparison, the highest (7.26 ± 0.37mg/kg of Z03 barrel plant soil available phosphorus content, n=10), in soil, the quantity of Z03 is also the highest [(5.43 ± 0.43) * 10
4cfu/g soil, n=10]; The effective pod number of Z03 individual plant (individual), single-strain grain number (grain), single-strain grain weight (g) and 100-grain weight (g) [are respectively 23.00 ± 1.66,47.60 ± 2.75,12.13 ± 0.82 g and 18.83 ± 0.50, (n=10)] compare with other group the highest, and compare with other group significant difference (
p<0.05).The effective pod number of Z03 individual plant, single-strain grain number, single-strain grain weight compare with control group with 100-grain weight average improve respectively 18.56%, 18.11%, 20.94% and the effective pod number of 6.8%(control group individual plant (individual), single-strain grain number (grain), single-strain grain weight (g) and 100-grain weight (g) be respectively 19.40 ± 1.12,40.30 ± 2.17,10.03 ± 0.19 and 17.63 ± 0.28, n=10).To the Molecular Identification of molten phosphorus fungi Z03 find with
penicilliumbelong to fungi similarity and reach 96%, sequential covering degree reaches 97%, so called after
penicilliumsp. XXR-G, the GenBank number of logging in KF572132, soybean tailored version biologic phosphorus fertilizer prepared by molten phosphorus fungi Z03, has obvious effect of increasing production.
Embodiment tri-
1) impact on mung bean output by the molten phosphorus fungi of 4 strain (being numbered Z01, Z02, Z03 and the Z04) test in embodiment mono-.
2) with ground phosphate rock fermented liquid, [ground phosphate rock fermented liquid, every liter contains: 10 g glucose, 10 g ground phosphate rock are (containing P
2o
526%), 5 g MgCl
26H2O, 0.25 g MgSO
47H
2o, 0.2 g KCl, 0.1 g (NH
4)
2sO
4)], enlarged culturing, cultivates 16 days for 28 ℃; Control group is not for adding the ground phosphate rock nutrient solution of molten phosphorus microorganism.
3) selecting cultivation bucket diameter is 25cm, dark 40cm, and bottom of the barrel borehole diameter 2cm is beneficial under excessive moisture and oozes.
4) cultivation bucket is put into the heatable adobe sleeping platform digging in advance, plane 5cm above ground level bucket on, then with the native hole tamping barrel with surrounding soil.
5) soil heatable adobe sleeping platform being dug out is removed macrobead with 2cm aperture particle size stainless (steel) wire, after repeatedly mixing, as cultivation matrix, 40kg soil barrelling having peace of mind is extremely cultivated to 4/5 height of bucket.
6) from planting site fetch earth (water content is 40-50%), with sieve aperture 6mm stainless (steel) wire, remove macrobead, the soil sieving is mixed with the mung bean cane of pulverizing, by soil: mung bean cane=8:1 (w/w), fully mix (mung bean cane water content is 40-50%), mixed soil is mixed with molten phosphorus fungi ground phosphate rock fermented liquid to (bacteria containing amount is 6.0 * 10
5cfu/g is native), or contrast ground phosphate rock nutrient solution mixes.The soil that contains phosphorus-solubilizing bacteria or contrast culture liquid is contained in to cultivation matrix top in cultivation bucket, reaches cultivation bucket top edges (cultivation bucket 1/5 position, top), during plantation, making soil moisture content is 60-75%, records planting soil available phosphorus content 9.23mg/kg.
7) mung bean seed (green No. 4 of Hubei Province) is planted in the soil of bucket internal upper part 1/5, after emerging, each barrel stays the seedling of 2 strain stalwartnesses.During results, control group soil available phosphorus content 4.61 ± 0.12mg/kg (n=10), with other phosphorus-solubilizing bacteria group comparison, the highest (5.23 ± 0.19mg/kg of Z04 barrel plant soil available phosphorus content, n=10), in soil, the quantity of Z04 is also the highest [(3.93 ± 0.27) * 10
4cfu/g soil, n=10], and significant difference (ρ <0.05).That Z04 single-strain legumen number (individual), Grain number per pod (grain) and single plant yield (g) are compared with other group is the highest (be respectively 20.80 ± 0.66,9.90 ± 0.38 and 8.24 ± 0.37, n=10), wherein single-strain legumen number compare with other group with single plant yield significant difference (
p<0.05).[control group single-strain legumen number (individual), Grain number per pod (grain) and single plant yield (g) are respectively 16.60 ± 0.81,8.50 ± 0.50 and 6.13 ± 0.38, n=10] compared and improved respectively 25.3%, 16.5% and 34.4% to Z04 single-strain legumen number (individual), Grain number per pod (grain) and single plant yield (g) average with control group.The ITS sequence of molten phosphorus fungi Z04 with registered
acremonium,
penicillium,
talaromyces and
trichocomaceaebelonging to fungi maximum similarity is 90%, called after Fungal sp. XXR-E, and the GenBank number of logging in KF572130, mung bean tailored version biologic phosphorus fertilizer prepared by molten phosphorus fungi Z04, has obvious effect of increasing production.
Claims (7)
1. a crop tailored version biologic phosphorus fertilizer, is characterized in that: this biologic phosphorus fertilizer is molten phosphorus fungi ground phosphate rock fermented liquid, and molten phosphorus fungi is
aspergillus nigerxXR-B1, during the GenBank number of logging in KF572126, makes corn tailored version biologic phosphorus fertilizer.
2. a crop tailored version biologic phosphorus fertilizer, is characterized in that: this biologic phosphorus fertilizer is molten phosphorus fungi ground phosphate rock fermented liquid, and molten phosphorus fungi is
penicilliumsp. XXR-G, during the GenBank number of logging in KF572132, makes soybean tailored version biologic phosphorus fertilizer.
3. a crop tailored version biologic phosphorus fertilizer, is characterized in that: this biologic phosphorus fertilizer is molten phosphorus fungi ground phosphate rock fermented liquid, and molten phosphorus fungi is Fungal sp. XXR-E, during the GenBank number of logging in KF572130, makes mung bean tailored version biologic phosphorus fertilizer.
4. a preparation method for crop tailored version biologic phosphorus fertilizer, is characterized in that, comprises the following steps:
1) gather soil sample, and this soil sample is inoculated in phosphorus-solubilizing bacteria screening culture medium after gradient dilution, at 25 ℃-30 ℃, cultivate 5-10 days, make the fungal colony edge in soil sample occur molten phosphorus circle, the fungal colony that produces molten phosphorus circle is forwarded in new phosphorus-solubilizing bacteria screening culture medium, at 25 ℃-30 ℃, cultivate 3-5 days, carry out obtaining molten phosphorus fungi after separation and purification;
2) the molten phosphorus fungi after separation and purification is placed in to ground phosphate rock fermented liquid, at 28 ℃ of-30 ℃ of bottom fermentations, cultivates 15-18 days, obtain molten phosphorus fungi ground phosphate rock fermented liquid, this molten phosphorus fungi ground phosphate rock fermented liquid is tailored version biologic phosphorus fertilizer.
5. crop tailored version biologic phosphorus fertilizer preparation method according to claim 1, is characterized in that: in step 1), by soil sample, through gradient dilution, be 10
-1, 10
-2, 10
-3.
6. crop tailored version biologic phosphorus fertilizer preparation method according to claim 1, is characterized in that: in step 1), phosphorus-solubilizing bacteria screening culture medium is that glucose, calcium phosphate, 6 aqueous magnesium chlorides, 7 water magnesium sulfates, Repone K, ammonium sulfate, agar be take to mass ratio as 15:5:5:0.25:0.2:0.1:15 mixing.
7. crop tailored version biologic phosphorus fertilizer preparation method according to claim 1, is characterized in that: step 2) in ground phosphate rock fermented liquid be by weight being 10:10:5:0.25:0.2:0.1 mixing by glucose, ground phosphate rock, 6 aqueous magnesium chlorides, 7 water magnesium sulfates, Repone K, ammonium sulfate.
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Cited By (1)
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Application publication date: 20141008 |