CN103141517B - Paenibacillus terrae biological agent and application thereof in agriculture - Google Patents
Paenibacillus terrae biological agent and application thereof in agriculture Download PDFInfo
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- 241000058353 Paenibacillus terrae Species 0.000 title claims abstract description 51
- 239000003124 biologic agent Substances 0.000 title abstract 9
- 239000007788 liquid Substances 0.000 claims abstract description 70
- 239000007787 solid Substances 0.000 claims abstract description 30
- 239000002689 soil Substances 0.000 claims description 124
- 241000193830 Bacillus <bacterium> Species 0.000 claims description 108
- 238000002360 preparation method Methods 0.000 claims description 70
- 244000068988 Glycine max Species 0.000 claims description 27
- 235000010469 Glycine max Nutrition 0.000 claims description 27
- 238000000855 fermentation Methods 0.000 claims description 16
- 230000004151 fermentation Effects 0.000 claims description 16
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 15
- 240000008042 Zea mays Species 0.000 claims description 15
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 15
- 235000015097 nutrients Nutrition 0.000 claims description 15
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 14
- 235000005822 corn Nutrition 0.000 claims description 14
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 claims description 13
- 238000010899 nucleation Methods 0.000 claims description 13
- 229930006000 Sucrose Natural products 0.000 claims description 10
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 10
- 244000144987 brood Species 0.000 claims description 10
- 239000005720 sucrose Substances 0.000 claims description 10
- 239000000843 powder Substances 0.000 claims description 9
- 238000004519 manufacturing process Methods 0.000 claims description 8
- 238000002156 mixing Methods 0.000 claims description 8
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- 235000013312 flour Nutrition 0.000 claims description 6
- 239000002054 inoculum Substances 0.000 claims description 6
- 230000001954 sterilising effect Effects 0.000 claims description 6
- 241000195940 Bryophyta Species 0.000 claims description 4
- 235000013339 cereals Nutrition 0.000 claims description 4
- 239000004615 ingredient Substances 0.000 claims description 4
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- 239000001888 Peptone Substances 0.000 claims description 3
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- 239000008120 corn starch Substances 0.000 claims description 3
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- 241000894006 Bacteria Species 0.000 description 19
- 229910052698 phosphorus Inorganic materials 0.000 description 19
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- 239000011574 phosphorus Substances 0.000 description 18
- 239000000047 product Substances 0.000 description 17
- 229910052700 potassium Inorganic materials 0.000 description 15
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 14
- 229910052757 nitrogen Inorganic materials 0.000 description 14
- 239000011591 potassium Substances 0.000 description 14
- 239000000126 substance Substances 0.000 description 13
- 238000002474 experimental method Methods 0.000 description 11
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- 235000009566 rice Nutrition 0.000 description 10
- 229910052710 silicon Inorganic materials 0.000 description 10
- 239000010703 silicon Substances 0.000 description 10
- DLHONNLASJQAHX-UHFFFAOYSA-N aluminum;potassium;oxygen(2-);silicon(4+) Chemical compound [O-2].[O-2].[O-2].[O-2].[O-2].[O-2].[O-2].[O-2].[Al+3].[Si+4].[Si+4].[Si+4].[K+] DLHONNLASJQAHX-UHFFFAOYSA-N 0.000 description 9
- 239000003795 chemical substances by application Substances 0.000 description 9
- 238000009335 monocropping Methods 0.000 description 9
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 8
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical group O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
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- 239000005696 Diammonium phosphate Substances 0.000 description 3
- 241000223221 Fusarium oxysporum Species 0.000 description 3
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- 238000009825 accumulation Methods 0.000 description 3
- 239000004202 carbamide Substances 0.000 description 3
- MNNHAPBLZZVQHP-UHFFFAOYSA-N diammonium hydrogen phosphate Chemical compound [NH4+].[NH4+].OP([O-])([O-])=O MNNHAPBLZZVQHP-UHFFFAOYSA-N 0.000 description 3
- 229910000388 diammonium phosphate Inorganic materials 0.000 description 3
- 235000019838 diammonium phosphate Nutrition 0.000 description 3
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- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 2
- 229930195725 Mannitol Natural products 0.000 description 2
- BPQQTUXANYXVAA-UHFFFAOYSA-N Orthosilicate Chemical compound [O-][Si]([O-])([O-])[O-] BPQQTUXANYXVAA-UHFFFAOYSA-N 0.000 description 2
- 241000179039 Paenibacillus Species 0.000 description 2
- 241001638069 Rigidoporus microporus Species 0.000 description 2
- 241000607479 Yersinia pestis Species 0.000 description 2
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- DUEPRVBVGDRKAG-UHFFFAOYSA-N carbofuran Chemical compound CNC(=O)OC1=CC=CC2=C1OC(C)(C)C2 DUEPRVBVGDRKAG-UHFFFAOYSA-N 0.000 description 2
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- VTNQPKFIQCLBDU-UHFFFAOYSA-N Acetochlor Chemical compound CCOCN(C(=O)CCl)C1=C(C)C=CC=C1CC VTNQPKFIQCLBDU-UHFFFAOYSA-N 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
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- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 1
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- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
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Abstract
The invention relates to a biological agent and application thereof in agriculture, particularly a Paenibacillus terrae biological agent and application thereof in agriculture. The Paenibacillus terrae biological agent contains Paenibacillus terrae NK3-4; and the Paenibacillus terrae biological agent is divided into a Paenibacillus terrae liquid biological agent and a Paenibacillus terrae solid biological agent. The Paenibacillus terrae liquid biological agent or Paenibacillus terrae solid biological agent can be applied. The invention is applicable to the field of agriculture.
Description
Technical field
The present invention relates to a kind of biologic product and the application in agricultural thereof.
Background technology
Nitrogen, phosphorus, potassium are the essential macronutrients of plant growth, and in plough horizon, content of tatal phosphorus is 0.1～2.0g/kg, and wherein Phos accounts for 15%～80% of full phosphorus total amount, and major part is the inorganic phosphorous compound of slightly solubility, are difficult to directly be utilized by plant; In plough horizon, full potassium content is 0.5～45g/kg, and wherein mineral potassium accounts for 90%～98% of full potassium, is difficult to directly be utilized by plant; The phosphate fertilizer of using in agricultural production and the availability of potash fertilizer only have 40%～60%, and remainder is retightened by soil, due to crop to low, the easy loss of its availability, cause serious waste and environmental pollution.
Because the mankind constantly increase the demand of agricultural products, cause crops continuous cropping to continue to occur, then produce continuous cropping obstacle.Continuous cropping meeting causes that soil physical and chemical property and biological character change, and in soil microbial community, beneficial microorganism growth is suppressed, and harmful microorganism is bred rapidly, and the natural equilibrium of edaphon is destroyed, and crop disease and insect occurs more and more serious.According to statistics, the underproduction that in Reducing production of continuous planting, disease causes accounts for more than 75%, and agricultural producer can only lean on traditional method of preventing and treating, continuing to increase dose controls with frequent medication, this not only can not solve plant disease problem, bring on the contrary serious side effect, caused the severe contamination of environment and agricultural product, reduced agricultural output and quality.
Summary of the invention
The present invention is in order to utilize the antagonism between microorganism to prevent and treat crop pest, reduce chemical pesticide consumption, alleviate the pollution of chemical pesticide to environment and agricultural product, utilize the nitrogen in microorganism fixed air, decompose mineral phosphorus, mineral K and slightly solubility silicon, and suppress soil-borne disease of crop; And a kind of soil series bacillus biologic product and the application in agricultural thereof are provided.
In soil of the present invention series bacillus biologic product, contain soil series bacillus (Paenibacillus terrae) NK3-4; Soil series bacillus biologic product is divided into soil series bacillus liquid bio preparation and soil series bacillus solid biologic preparation.
Use above-mentioned soil series bacillus liquid bio preparation or soil series bacillus solid biologic preparation, realized the application of soil series bacillus biologic product in agricultural.
Biomaterial soil series bacillus (Paenibacillus terrae) NK3-4 using in the present invention belongs to series bacillus and belongs to (Paenibacillus Ash, Priest & Collin, 1994), be deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation date is on December 17th, 2012, and preserving number is CGMCC No.7011.
Soil of the present invention series bacillus biologic product, in the situation that reducing applying quantity of chemical fertilizer, still can make paddy rice, corn, Soybean and Other Crops volume increase, and to be particularly used in conjunction with increasing production of rice amplitude more remarkable with siliceous fertilizer.
Soil of the present invention series bacillus biologic product is inhibited to pathogenic Fusarium oxysporum, can utilize this characteristic to prevent and treat crop pest, reduces chemical pesticide consumption, alleviates the pollution of chemical pesticide to environment and agricultural product.Soil of the present invention series bacillus biologic product is Antifungi soil-borne disease effectively, and for secondary soybean soil, soybean seedling root rot disease index declines 23 percentage points; Can by the mineral phosphorus in soil and fertilizer, slightly solubility silicon and mineral K rapid conversion, be rapid available phosphorus, quick-acting silicon and the available potassium that crop is easy to absorb; In soil, produce several physiological active substances, promote plant growth, improve the output of crop, during particularly with siliceous fertilizer compounding application, can significantly improve crop yield.
Chemical fertilizer, excessive the using of agricultural chemicals are the important sources polluting in agricultural production, and the environmental problem and the food-safety problem that cause are thus on the rise.The application of soil of the present invention series bacillus biologic product in agricultural can reduce the usage amount of chemical fertilizer and agricultural chemicals, have noresidue, free from environmental pollution, do not damage the advantages such as people and animals Jiankang, be the another new highlight of ecological agriculture development.
Soil series bacillus (Paenibacillus terrae) NK3-4 belongs to series bacillus and belongs to (Paenibacillus Ash, Priest & Collin, 1994), be deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC), preservation address is Datun Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, preservation date is on December 17th, 2012, preserving number is CGMCC No.7011.
Accompanying drawing explanation
Fig. 1 is the microscopic examination figure of soil series bacillus (Paenibacillus terrae) NK3-4.
Fig. 2 is the colonial morphology figure of soil series bacillus (Paenibacillus terrae) NK3-4 on silicate sucrose nitrogen-free agar flat board.
Embodiment
Technical solution of the present invention is not limited to following cited embodiment, also comprises any combination between each embodiment.
Embodiment one: contain soil series bacillus (Paenibacillus terrae) NK3-4 in present embodiment soil series bacillus biologic product; Soil series bacillus biologic product is divided into soil series bacillus liquid bio preparation and soil series bacillus solid biologic preparation.
Soil series bacillus in present embodiment (Paenibacillus terrae) NK3-4 is gram-positive bacteria, thalline is shaft-like, 1.0～1.2 μ m * 3～4 μ m, can form brood cell, while forming brood cell, thalline near-end expands, cyst obviously expands, and brood cell is oval, is positioned at cyst one end (as shown in Figure 1).Soil series bacillus (Paenibacillus terrae) NK3-4 is on silicate sucrose nitrogen-free agar flat board, cultivate and within 5～7 days, form the circular bacterium colony that diameter is 1～2mm left and right, single bacterium colony median rise, edge is irregular, surface wettability, translucent, lawn is dense thick, easily provokes (as shown in Figure 2).Soil series bacillus (Paenibacillus terrae) NK3-4 bacterium colony on potassium feldspar dusty yeast medium is creamy white, half glass pearl, circular bacterium colony, lawn amount, gluing more, be cultured to later stage colony edge irregular, growth-gen can grow to medium inside, and medium is split.
Soil series bacillus (Paenibacillus terrae) NK3-4 analyzes by 16S rDNA sequence alignment, the most approaching with Paenibacillus terrae AM141 bacterial strain (accession number is AF391124) affiliation, similitude is 99%, but soil series bacillus (Paenibacillus terrae) NK3-4 is again from Paenibacillus terrae AM141, to compare existence significantly different.Soil series bacillus (Paenibacillus terrae) NK3-4 is Gram-positive, and thalline length is 1.0～1.2 μ m * 3～4 μ m, can not utilize mannitol, in 2% NaCl, does not grow; And Paenibacillus terrae AM141 Gram’s staining is variable, thalline length is 1.3～1.8 μ m * 4～7 μ m, can utilize mannitol, in 2% NaCl, grows.
Soil series bacillus in present embodiment (Paenibacillus terrae) NK3-4 Physiological-biochemical Characters and utilize situation as shown in table 1 to carbon and nitrogen sources.
Table 1
Soil series bacillus (Paenibacillus terrae) NK3-4 has the nitrogen in fixed air, decomposes the function of mineral phosphorus (Phos), mineral K and slightly solubility silicon.
Soil series bacillus (Paenibacillus terrae) NK3-4 decomposes mineral, discharges potassium, silicon, phosphorus, the nitrogen in fixed air:
Soil series bacillus (Paenibacillus terrae) NK3-4 bacterial strain can think that phosphate ore is to grow on the solid plate medium in unique phosphorus source, containing 1% Ca
3(PO
4)
2phosphorus decomposing culture fluid in cultivate after 8d, the concentration of measuring rapid available phosphorus in culture fluid with molybdenum antimony resistance colorimetric method is 57.6～70.5mg/L.
Soil series bacillus (Paenibacillus terrae) NK3-4 bacterial strain shows the ability of stronger decomposition silicate mineral under shaking flask condition.Take feldspar in powder as substrate, cultivate after 24 hours, the concentration of medium effective K is 127.3 mg/L, thalline effective K concentration is 22.0 mg/L, culture fluid effective K concentration does not more add potassium feldspar and processes high 15.0 mg/L of contrast, and fixing potassium does not more add high 4.8 mg/L of processing control group of potassium feldspar in soil series bacillus (Paenibacillus terrae) NK3-4 thalline.
Soil series bacillus (Paenibacillus terrae) NK3-4 bacterial strain is when destroying silicate mineral, also discharge element silicon, take potassium feldspar as substrate, and the concentration that connects the silicon in bacterium processing filtrate is 193.0～226.2 mg/L, and the control group of inoculating inactivated bacteria processing is high by 131.7%.
Soil series bacillus (Paenibacillus terrae) NK3-4 bacterial strain is cultivated 15d at A Xubei (Ashby) in without nitrogen culture fluid, and in bacterium liquid, quick-acting nitrogen contents can reach 8.1～11.6 mg/L, and the control group of processing than inoculation inactivated bacteria is high by 368.1%.
Embodiment two: the difference of present embodiment and embodiment one is: the cultivation and fermentation liquid of soil series bacillus liquid bio preparation for containing soil series bacillus (Paenibacillus terrae) NK3-4.Other step and parameter are identical with embodiment one.
Soil series bacillus liquid bio preparation has obvious inhibitory action to pathogenic Fusarium oxysporum.On the PDA medium of inoculation Fusarium oxysporum, drip soil series bacillus liquid bio preparation and carry out plate face-off cultivation, occur obvious inhibition zone.
Brood cell's quantity 1 * 10 in present embodiment soil series bacillus liquid bio preparation
9～2 * 10
9individual/ml, field crop is used soil series bacillus liquid bio preparation 3.5～9 L/hm of the present invention
2, can make crop yield more than 8%, with siliceous fertilizer compounding application, can make increasing production of rice 16%.
Embodiment three: present embodiment with embodiment one or two difference is: soil series bacillus solid biologic preparation is mixed by major ingredient soil series bacillus liquid bio preparation and the peat composed of rotten mosses, then extruder grain obtains soil series bacillus solid biologic preparation.Other step and parameter are identical with embodiment one or two.
The use of present embodiment soil series bacillus solid biologic preparation can be not only increasing crop yield, more has the effect that reduces fertilizer application amount.
Embodiment four: present embodiment and embodiment one, two or threes' difference is: soil series bacillus liquid bio preparation is prepared according to the following steps:
A, by soil series bacillus (Paenibacillus terrae) NK3-4 kind to liquid nutrient medium, under 180 r/min conditions, cultivate 13h, then by 0.5% inoculum concentration, be inoculated in the seeding tank that liquid nutrient medium is housed, in seeding tank, the volume of liquid nutrient medium is 70% of seeding tank volume, at 28～30 ℃, mixing speed, is that 220r/min and filtrated air intake are to cultivate 11h under the condition of 1 ︰ 0.8; Every liter of liquid nutrient medium contains 10g sucrose, 0.2g K
2hPO
4, 0.2 g MgSO
4, 0.2 g NaCl, 5.0g CaCO
3, 1.0g CaSO
4, 2.0g dusty yeast, 5.0g starch, 1.0g (NH
4) SO
4, pH value 7.2～7.5;
B, through the soil of seed tank culture series bacillus (Paenibacillus terrae) NK3-4, by 5% inoculum concentration, be inoculated in the production tank that fermentation medium is housed, produce the volume of fermentation medium in tank for producing 70% of tank volume, in fermentation medium per ton, contain 1.2kg sucrose, 2.6kg corn starch, 1.2kg peptone, 0.84kg dried yeast, 0.24kg magnesium sulfate, 0.6kg CaCO
3, 1.2kg K
2hPO
4, 6.3kg soybean cake powder, 18.0kg corn flour, 0.9kg yeast, 0.15L defoamer and 1.35kg soya-bean oil, pH value 7.2～7.5 after sterilizing; Then at 28～30 ℃, mixing speed, be that 250r/min and filtrated air intake are that in being cultured to soil series bacillus (Paenibacillus terrae) NK3-4 zymotic fluid under the condition of 1 ︰ 0.8,95% above thalline produces brood cell, and brood cell's quantity surpasses 1,000,000,000/ml, obtains soil series bacillus liquid bio preparation.Other step and parameter are identical with embodiment one, two or three.
Present embodiment is according to seeding tank and the difference of producing tank radius, and present embodiment has been selected different fermentation mixing speeds.
In present embodiment steps A seeding tank must be first with steam sterilizing and be cooled to 28～30 ℃, filling liquid medium then.In step B, produce tank in advance with steam sterilizing and be cooled to 28～30 ℃, then inject fermentation medium.
Embodiment five: the difference of present embodiment and embodiment four is: in step B, beancake powder and corn flour add the 24h that soaks and ferment after dried yeast, and then grind and add fermentation medium.Other step and parameter are identical with embodiment four.
Embodiment six: present embodiment and embodiment three, four or fives' difference is: in soil series bacillus solid biologic preparation, the mass ratio of soil series bacillus liquid bio preparation and the peat composed of rotten mosses is 1 ︰ 10, and add the white clay of soil series bacillus solid biologic preparation gross mass 30%～35% and 10%～15% humus as auxiliary material.Other step and parameter are identical with embodiment three, four or five.
Living bacteria count >=0.2 hundred million/g in present embodiment soil series bacillus solid biologic preparation, the content of organic matter >=40%.Field crop is used present embodiment soil series bacillus solid biologic preparation 30～75kg/hm
2, can make crop yield more than 8%.
Embodiment seven: present embodiment is used soil series bacillus liquid bio preparation or soil series bacillus solid biologic preparation, has realized the application of soil series bacillus biologic product in agricultural.
Embodiment eight: the difference of present embodiment and embodiment seven is: soil series bacillus liquid bio preparation and crop seeds are uniformly mixed, then sowing; Every kilogram of crop seeds and 10～15ml soil series bacillus liquid bio preparation are puddled evenly.Other step and parameter are identical with embodiment seven.
Embodiment nine: present embodiment with the difference of embodiment seven is: fertilising after soil series bacillus liquid bio preparation mixes with chemical fertilizer, as base manure, on paddy rice, use, soil series bacillus liquid bio preparation consumption is 3.5～9.0L/hm
2; Or foliage-spray soil series bacillus liquid bio preparation, soil series bacillus liquid bio preparation consumption is 1.2～2.0L/hm
2.Other step and parameter are identical with embodiment seven.
Embodiment ten: present embodiment with the difference of embodiment seven is: use after soil series bacillus solid biologic preparation mixes with chemical fertilizer, soil series bacillus solid biologic preparation consumption is 30～75kg/hm
2.Other step and parameter are identical with embodiment seven.
Embodiment 1
Picking soil series bacillus (Paenibacillus terrae) NK3-4 lawn is inoculated in potassium feldspar liquid nutrient medium, and (potassium feldspar liquid nutrient medium is by 10g sucrose, 0.2g K
2hPO
4, 0.2g MgSO
47H
2o, 0.2g NaCl, 5.0g CaCO
3, 1.0g feldspar in powder, 0.1g CaSO
4, 2.0g dusty yeast and 1L distilled water forms, pH value 7.2), (control group medium is by 10g sucrose, 0.2g K in addition soil series bacillus (Paenibacillus terrae) NK3-4 to be inoculated in not to the control group medium of K-feldspar powder
2hPO
4, 0.2g MgSO
47H
2o, 0.2g NaCl, 5.0g CaCO
3, 0.1g CaSO
4, 2.0g dusty yeast and 1L distilled water forms, pH value is 7.2), with the potassium feldspar liquid nutrient medium that do not connect bacterium, as blank, test and establish 4 repetitions, under 30 ℃, 200r/min condition, cultivate 24h, standby.
The bacterium liquid of getting above-mentioned cultivation 24h dilutes 10 times, by flame spectrometry, survey the concentration of potassium in medium, separately get bacterium liquid 5ml in 100 ℃ of water-bath heating 30min, so that bacterial cell disruption, discharge potassium, cooling after, dilute 10 times, measure liquid effective K concentration (first measure medium and thalline potassium concn with, this value deducts the value of direct mensuration, is the concentration of thalline potassium in every liter of liquid).Result shows, in the concentration of inoculating the potassium feldspar medium effective K of soil series bacillus (Paenibacillus terrae) NK3-4, be 127.3mg/L, thalline effective K concentration is 22.0mg/L, and the concentration of the control group medium effective K of K-feldspar is not 112.3mg/L, thalline effective K concentration is 17.3mg/L.Exceed respectively 15.0mg/L and 4.8mg/L.
Get 10 times of filtrations afterwards of bacterium liquid dilution of above-mentioned potassium feldspar medium culture, with molybdenum blue spectrophotometric method, measure the concentration of quick-acting silicon in filtrate, the concentration that result shows to inoculate silicon in the filtrate of soil series bacillus (Paenibacillus terrae) NK3-4 bacterium is 193.0～226.2mg/L, the concentration that does not connect silicon in bacterium blank group is 89.0mg/L, higher by 131.7% than the blank group of not inoculating.
Soil series bacillus (Paenibacillus terrae) NK3-4 bacterial strain is cultivated 30 ℃ of 15d(condition of culture, 200r/min at A Xubei (Ashby) in without nitrogen culture fluid), in bacterium liquid, quick-acting nitrogen contents can reach 8.1～11.6mg/L, the blank group available nitrogen content of not inoculating is 2.06mg/L, and NK3-4 is higher by 368.1% than the blank group of not inoculating for inoculation soil series bacillus (Paenibacillus terrae).
Soil series bacillus (Paenibacillus terrae) NK3-4 is inoculated in phosphorus decomposing culture fluid to (every liter of phosphorus decomposing culture fluid contains glucose 10.0g, NaCl0.3g, MnSO
44H
2o 0.03g, (NH
4)
2sO
40.5g, KCl 0.3g, FeSO
47H
2o0.03g and Ca
3(PO
4)
210.0g, pH value 7.2), not connect bacterium in contrast, repeat for 4 times, under 30 ℃, 200r/min condition, cultivate after 8d, measure the concentration of rapid available phosphorus in culture fluid with molybdenum blue colorimetric method, in the medium of inoculation soil series bacillus (Paenibacillus terrae) NK3-4, the concentration of rapid available phosphorus is 57.6～70.5mg/L, do not connect in bacterium control group as 13.3mg/L, the former is 4.8 times of the latter.
Embodiment 2
It is that 0.1% liquor natrii hypochloritis sterilizes 3 minutes that selected soya seeds is put into concentration, then take out clean with aseptic water washing rapidly, move into again in the aseptic flat board of a sterilizing circle filter paper, 15/plate, add soil series bacillus liquid bio preparation (all unitedly calling below NK3-4 liquid bio preparation) 10ml, and to add 10ml sterile water as a control group, repeat for 9 times, within the 3rd day, survey germination vigor, within the 6th day, survey germination rate.
Experimental result is as shown in table 2, and NK3-4 liquid bio preparation can make soya seeds germination rate improve 4.4 percentage points.
Experiment conclusion: NK3-4 liquid bio preparation can be done soya seeds presowing treatment.
Table 2
Embodiment 3
For examination soil, be continuous cropping soil (5 years Soybean Fields of continuous cropping) and main crop soil (front stubble is the Soybean Field of corn), in the soyabean field of front stubble for corn, sickle-like bacteria spore do not detected.Get continuous cropping soil and sieve with main crop folk song is dry, be loaded in 2.2L basin alms bowl, every alms bowl fills native 2.35kg.Experiment is divided into 3 groups, the 1st group: before soya seeds sowing by seed coat agent (effective ingredient is 15% carbendazim, 10% tmtd and 10% carbofuran) by the weight ratio Dressing of seed coat agent and soya seeds 1 ︰ 100; The 2nd group: before soya seeds sowing, use NK3-4 liquid bio preparation in the ratio Dressing of 1000 grams of soya seeds of 10ml NK3-4 liquid bio Ji ︰ processed; The 3rd group: using in proportion clear water Dressing as blank group.6 seeds of every alms bowl sowing, field planting after emerging, every alms bowl 2 strains, establish 4 repetitions.In soybean the 3rd compound leaf and the 5th compound leaf, measure plant height, dry-matter accumulation amount, dross situation, root rot disease index period.
Experimental result is as shown in table 3, by NK3-4 liquid bio preparation and seed coat agent, all can reduce dry matter accumulation of soybean amount and disease index in main crop soil, and wherein NK3-4 liquid bio preparation has obviously increased root nodule numbers and weight.In continuous cropping soil, NK3-4 liquid bio preparation and seed coat agent are to beans seedling dry-matter accumulation amount, root nodule dry weight and quantity all higher than blank group, and disease index is all lower than blank group.The result of use of NK3-4 liquid bio preparation is slightly worse than seed coat agent 3 compound leaf phases, 5 compound leaf phase effects, is better than seed coat agent.Experiment shows to apply the effect lasting stability of NK3-4 liquid bio preparation.
Table 3
Embodiment 4
Experiment is arranged on soybean second crop soil (4 years Soybean Fields of continuous cropping), establishes conventional fertilizer application and conventional fertilizer application and uses two groups of NK3-4 liquid bio preparation Dressings.Soybean varieties is rich 16 for cultivating, and 100-grain weight is 18g, hectare 360,000 strains of keeping a full stand of seedings.Community experiment, establishes 3 repetitions, and random district group is arranged, and community area is 16.25m
2.Conventional fertilizer application: urea 40kg/hm
2, diammonium phosphate 150kg/hm
2, potassium chloride 40kg/hm
2; Conventional fertilizer application is also dressed seed with NK3-4 liquid bio preparation, and NK3-4 liquid bio preparation consumption is 10ml/kg seed.
With NK3-4 liquid bio preparation Dressing, mu Yield of Soybean amount reaches 2680.5kg/hm
2, than conventional fertilizer application (conventional fertilizer application per mu yield 2424.0kg/hm
2) high by 10.6%.
Embodiment 5
Carry out in experiment field, land-reclaimable academy of sciences rice research institute garden, 2012 Nian Heilongjiang Province, soil organic matter 20.5g/kg, full nitrogen 1.1g/kg, rapid available phosphorus 10.2mg/kg, available potassium 218mg/kg, pH value 6.5.Community area 24m
2, line-spacing 25cm, cave is apart from 10cm.
Experiment arranges 9 experimental group, 3 repetitions, and for examination rice varieties, for cultivating rice 10, each experimental group fertilising situation is as shown in table 4.
Table 4
Field management measure is all undertaken by locality plantation conventional method, samples, species test during rice harves, in each community, chooses 3m
2harvesting is surveyed and is produced.Species test and survey to produce result as shown in table 5, what conventional fertilizer application enriched siliceous fertilizer and used that NK3-4 liquid bio preparation of the present invention processes can obviously improve rice yield constituent element, and paddy rice average yield reaches 10008.3kg/hm
2, normal fertilising volume increase 16.4%, more only enriches siliceous fertilizer volume increase 10.2%.
Table 5
Embodiment 6
For examination seed coat agent effective ingredient, be 15% carbendazim, 10% tmtd, 10% carbofuran; (executing can rich board: N13%, P for commercial anti continuous and alternative-year composite fertilizer special
2o
525%, K
2o10%, micro-fertilizer such as molybdenum); Urea (N:46%), diammonium phosphate (P
2o
5: 46%, N:18%), potassium chloride (K
2o:60%), soybean varieties is rich 16 for cultivating, and 100-grain weight is 18 grams, hectare 360,000 strains of keeping a full stand of seedings.
Sample plot soil types is black earth, soybean continuous cropping 6 years, the content of organic matter is 3.9%, full nitrogen 1.522g/kg, rapid available phosphorus 43mg/kg) and available potassium 170mg/kg, pH value 6.8.
4 experimental group are established in experiment, repeat community area 16.25m for 4 times
2, soil series bacillus solid biologic preparation (all unitedly calling below NK3-4 solid biologic preparation) per hectare consumption is 45kg, after mixing, so that base manure form is disposable, applies with chemical fertilizer.Experimental group 1:NK3-4 solid biologic preparation+80% conventional fertilizer application; Experimental group 2: executing can rich board anti continuous and alternative-year composite fertilizer special per hectare 300kg; Experimental group 3:1.3% seed coat agent seed dressing+conventional fertilizer application; Experimental group 4: conventional fertilizer application.Conventional fertilizer application: per hectare urea 40kg, diammonium phosphate 150kg and potassium chloride 40kg.
Single-strain grain number, the grain of using as can be seen from Table 6 NK3-4 solid biologic preparation (experimental group 1) are heavy the highest, and individual plant pod is heavy, stem is heavy also in prostatitis.Experimental group 1 output is the highest, than conventional fertilizer application (experimental group 4) volume increase 10.06%, than seed coat agent (experimental group 3) volume increase 7.3%.When using the volume increase of NK3-4 solid biologic preparation, also reduced by 20% applying quantity of chemical fertilizer.
Table 6
Embodiment 7
Experiment is located in land-reclaimable academy of sciences's crop institute's corn test block, Heilongjiang Province, and soil is meadow chernozemic soil.Soil plough horizon plinth nutrient content is organic matter 3.26%, hydrolyzable nitrogen 46.1%, rapid available phosphorus 150.3mg/kg, available potassium 229.6mg/kg, pH value 6.9.Front stubble is corn, autumn subsoiling ridging.During sowing, soil moisture is not enough, and after Maize at Seedling Stage, jointing, rainfall is plentiful.For trying corn variety: four morning No. 11.
The experiment of employing community, 6 row districts, the long 8m of row, repeats for 3 times, and random district group is arranged, community area 31.2m
2.Experiment is established 10 groups, and fertilising situation is as shown in table 7.
Table 7
Sowing on May 7, artificial ditching-fertilizing, equidistant program request, density 60,000 strains/hm
2.Before broadcasting rear seedling, with Acetochlor 1500ml+ match gram 400g, carry out enclosed weeding, 4～5 leaf phase of corn subsoiling.September 26～28 gathered in the crops.
Experimental result is as shown in table 8, significant difference between NK3-4 solid biologic preparation and control group (experimental group 10), NK3-4 solid biologic preparation makes seed manure and seed coordination, plant under, plant and enclose that to use a difference not remarkable, and increase production more than 7% compared with conventional fertilizer application contrast, NK3-4 solid biologic preparation is made seed manure, leaf fertilizer, a significant difference that topdresses, best by effect of increasing production as seed-fertilizer applying; NK3-4 solid biologic preparation is as seed manure, and amount of application is 30kg/hm
2, 45kg/hm
2, 75kg/hm
2between difference not remarkable, and all compared with conventional fertilizer application contrast volume increase 10%.Topdressing normally use in the situation that, by corn conventional fertilizer application amount, to calculate: soil series bacillus biosolids preparation 25%+ chemical fertilizer 75% is done seed manure, output is executed and is improved 3.9%～8.2% than routine.
Table 8
Claims (5)
1. soil series bacillus biologic product, it is characterized in that soil series bacillus biologic product is soil series bacillus liquid bio preparation, the cultivation and fermentation liquid of soil series bacillus liquid bio preparation for containing soil series bacillus (Paenibacillus terrae) NK3-4; Soil series bacillus liquid bio preparation is prepared according to the following steps:
A, soil series bacillus (Paenibacillus terrae) NK3-4 is seeded to liquid nutrient medium, under 180r/min condition, cultivate 13h, then by 0.5% inoculum concentration, be inoculated in the seeding tank that liquid nutrient medium is housed, in seeding tank, the volume of liquid nutrient medium is 70% of seeding tank volume, at 28～30 ℃, mixing speed, is to cultivate 11h under 220r/min and the filtrated air intake condition that is 1:0.8; Every liter of liquid nutrient medium contains 10g sucrose, 0.2g K
2hPO
4, 0.2g MgSO
4, 0.2gNaCl, 5.0g CaCO
3, 1.0g CaSO
4, 2.0g dusty yeast, 5.0g starch, 1.0g (NH
4) SO
4, pH value 7.2～7.5;
B, through the soil of seed tank culture series bacillus (Paenibacillus terrae) NK3-4, by 5% inoculum concentration, be inoculated in the production tank that fermentation medium is housed, produce the volume of fermentation medium in tank for producing 70% of tank volume, in fermentation medium per ton, contain 1.2kg sucrose, 2.6kg corn starch, 1.2kg peptone, 0.84kg dried yeast, 0.24kg magnesium sulfate, 0.6kg CaCO
3, 1.2kg K
2hPO
4, 6.3kg soybean cake powder, 18.0kg corn flour, 0.9kg yeast, 0.15L defoamer and 1.35kg soya-bean oil, pH value 7.2～7.5 after sterilizing; Then at 28～30 ℃, mixing speed, be that in being cultured to soil series bacillus (Paenibacillus terrae) NK3-4 zymotic fluid under 250r/min and the filtrated air intake condition that is 1:0.8,95% above thalline produces brood cell, and brood cell's quantity surpasses 1,000,000,000/ml, obtains soil series bacillus liquid bio preparation.
2. soil according to claim 1 series bacillus biologic product, is characterized in that in step B that beancake powder and corn flour add the 24h that soaks and ferment after dried yeast, and then grinds and add fermentation medium.
3. soil series bacillus biologic product, it is characterized in that soil series bacillus biologic product is soil series bacillus solid biologic preparation, soil series bacillus solid biologic preparation is mixed by major ingredient soil series bacillus liquid bio preparation and the peat composed of rotten mosses, then extruder grain obtains soil series bacillus solid biologic preparation; Soil series bacillus liquid bio preparation is prepared according to the following steps:
A, soil series bacillus (Paenibacillus terrae) NK3-4 is seeded to liquid nutrient medium, under 180r/min condition, cultivate 13h, then by 0.5% inoculum concentration, be inoculated in the seeding tank that liquid nutrient medium is housed, in seeding tank, the volume of liquid nutrient medium is 70% of seeding tank volume, at 28～30 ℃, mixing speed, is to cultivate 11h under 220r/min and the filtrated air intake condition that is 1:0.8; Every liter of liquid nutrient medium contains 10g sucrose, 0.2g K
2hPO
4, 0.2g MgSO
4, 0.2gNaCl, 5.0g CaCO
3, 1.0g CaSO
4, 2.0g dusty yeast, 5.0g starch, 1.0g (NH
4) SO
4, pH value 7.2～7.5;
B, through the soil of seed tank culture series bacillus (Paenibacillus terrae) NK3-4, by 5% inoculum concentration, be inoculated in the production tank that fermentation medium is housed, produce the volume of fermentation medium in tank for producing 70% of tank volume, in fermentation medium per ton, contain 1.2kg sucrose, 2.6kg corn starch, 1.2kg peptone, 0.84kg dried yeast, 0.24kg magnesium sulfate, 0.6kg CaCO
3, 1.2kg K
2hPO
4, 6.3kg soybean cake powder, 18.0kg corn flour, 0.9kg yeast, 0.15L defoamer and 1.35kg soya-bean oil, pH value 7.2～7.5 after sterilizing; Then at 28～30 ℃, mixing speed, be that in being cultured to soil series bacillus (Paenibacillus terrae) NK3-4 zymotic fluid under 250r/min and the filtrated air intake condition that is 1:0.8,95% above thalline produces brood cell, and brood cell's quantity surpasses 1,000,000,000/ml, obtains soil series bacillus liquid bio preparation.
4. soil according to claim 3 series bacillus biologic product, is characterized in that in step B that beancake powder and corn flour add the 24h that soaks and ferment after dried yeast, and then grinds and add fermentation medium.
5. according to the soil series bacillus biologic product described in claim 3 or 4, it is characterized in that in soil series bacillus solid biologic preparation, the mass ratio of soil series bacillus liquid bio preparation and the peat composed of rotten mosses is 1:10, and add the white clay of soil series bacillus solid biologic preparation gross mass 30%～35% and 10%～15% humus as auxiliary material.
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