CN103146622B - Paenibacillus terrae strain - Google Patents
Paenibacillus terrae strain Download PDFInfo
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- CN103146622B CN103146622B CN201310097450.2A CN201310097450A CN103146622B CN 103146622 B CN103146622 B CN 103146622B CN 201310097450 A CN201310097450 A CN 201310097450A CN 103146622 B CN103146622 B CN 103146622B
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- paenibacillus terrae
- series bacillus
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- terrae
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- 239000011591 potassium Substances 0.000 abstract description 26
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Abstract
The invention relates to paenibacillus elgii, particularly a Paenibacillus terrae strain. The Paenibacillus terrae NK3-4 is collected at China General Microbiological Culture Collection Center, and the collection number is CGMCC No.7011. The NK3-4 has the functions of fixing nitrogen in air, decomposing mineral phosphorus, mineral potassium and insoluble silicon, and inhibiting phytopathogens. The Paenibacillus terrae NK3-4 can still increase the yield of rice, corn, soybean and other crops under the conditions of reducing the application amount of the fertilizer, and especially has more obvious yield increase amplitude when being matched with a silicon fertilizer. The invention is applicable to the field of agriculture.
Description
Technical field
The present invention relates to a kind of series bacillus.
Background technology
Class Bacillus (Paenibacillus Ash, Priest & Collin, 1994) belongs to Firmicutes, bacillus guiding principle, bacillus order, class Bacillaceae.It is shaft-like that cell is, and Gram-positive, feminine gender or variable, move with peritrichous.In cyst, have oval gemma expanding, on nutrient agar medium without soluble pigment.Amphimicrobian or strictly aerobic.At least recorded at present 39 kinds, class bacillus is widely distributed at occurring in nature.
The research of known class bacillus is found to it has high using value.For example, Paenibacillus polymyxa (P.polymyxa) can be prevented and treated the multiple harmful microorganisms such as pathogenetic bacteria, pathogenic fungi, root knot nematode, can secrete multiple peptide class antimicrobial substance (as polymyxin, Totazina, circulin, jolipeptin, circulin, gatavalin, kill fusanin etc.), can directly act on again bacterium thalline, and can induce host plant to produce disease resistance, can also, by fixed nitrogen and dissolved organic phosphorus for host plant provides nutrition, promote plant strain growth.Fixed nitrogen class bacillus (P.azotofixans) has obvious restraining effect to excellent bacillus (Corynebacterium fimi), and heavy metal and microbiotic are had to stronger tolerance.And also found in recent years that some possessed the class bacillus novel species of new function, the calendar year 2001s such as Aguilera have found that a strain has the novel species P.s jamilae that produces exocellular polysaccharide function in the straw compost of olive squeezing waste material composting; Daane equals at sabkha plant rhizosphere, to find can the degrade novel species P.naphthalenovorans of naphthalene of a strain in 2002; Yoon etc. have found that two strains can produce novel species P.kribbensis and the P.terrae of the flocculation agent of efficient flocculating marine alga for 2003; Horn etc. have found that a strain can produce N for 2005 in earthworm internal organ
2the novel species P.anaericanus of O.
Soil class bacillus (P.s terrae) is the novel species of finding in India in 2003, having at present 3 soil class bacterial strain of bacillus to be reported as function stem, is respectively Paenibacillus terrae AM141 bacterial strain, HPL-003 bacterial strain and MH72 bacterial strain.Paenibacillus terrae AM141 bacterial strain can be used as the flocculation agent of algae results, and HPL-003 bacterial strain can produce zytase, and MH72 bacterial strain can produce N
2o.
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 inorganic phosphorus accounts for 15%～80% of full phosphorus total amount, and wherein major part is the inorganic phosphorous compound of insoluble, 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 agriculture production and the utilization ratio of potash fertilizer only have 40%～60%, and rest part is retightened by soil, due to crop to low, the easy loss of its utilization ratio, cause serious waste and environmental pollution.
Summary of the invention
The invention provides a strain soil series bacillus
Soil of the present invention series bacillus (Paenibacillus terrae) NK3-4 bacterial strain, is deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and preserving number is CGMCC No.7011.
Soil of the present invention series bacillus (Paenibacillus terrae) NK3-4 belongs to series bacillus and belongs to (PaenibacillusAsh, 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 series bacillus (Paenibacillus terrae) NK3-4 is gram-positive microorganism, and 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, and cyst obviously expands, brood cell's ellipse, 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 felspar sand yeast powder substratum is creamy white, half glass pearl, circular bacterium colony, lawn amount, gluing more, be cultured to later stage colony edge irregular, grower can grow to substratum inside, and substratum is split.
Soil series bacillus (Paenibacillus terrae) NK3-4 analyzes by 16S rDNA sequence alignment, and the most approaching with Paenibacillus terrae AM141 bacterial strain (accession number is AF391124) sibship, similarity is 99%.The 16S rDNA of soil series bacillus (Paenibacillus terrae) NK3-4 is as shown in SEQ ID NO:1.
To compare existence significantly different from Paenibacillus terrae AM141 again for soil series bacillus (Paenibacillus terrae) NK3-4.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 N.F,USP MANNITOL, in 2%NaCl, does not grow; And Paenibacillus terrae AM141 gramstaining is variable, thalline length is 1.3～1.8 μ m × 4～7 μ m, can utilize N.F,USP MANNITOL, in 2%NaCl, grows.Therefore, judge the new bacterial strain of a strain that soil series bacillus (Paenibacillus terrae) NK3-4 is soil series bacillus kind.
Soil of the present invention series bacillus (Paenibacillus terrae) NK3-4 gramstaining is positive, and indole reaction is negative, and M.R test is negative, and V.P test is positive; Well-grown on silicate sucrose nitrogen-free agar.Soil series bacillus (Paenibacillus terrae) NK3-4 bacterial strain can well utilize glucose, sucrose, lactose, Xylo-Mucine, humic acid etc. as carbon source, and energy hydrolyzed starch, does not utilize N.F,USP MANNITOL; While utilizing dextrose plus saccharose, produce sour aerogenesis, produce not aerogenesis of acid while utilizing lactose, Xylo-Mucine, humic acid, after general cultivation, substrate pH value reduces by 0.8～1.5.Soil series bacillus (Paenibacillus terrae) NK3-4 can utilize the nitrogen in most of amino acid, can make gelatine liquefication, milk is peptonized, and in 1%NaCl, grows, and in 2%NaCl, does not grow; Optimum growth temperature is 30～32 ℃, and growth optimum pH is 6.5～7.5.Soil series bacillus (Paenibacillus terrae) NK3-4 Physiological-biochemical Characters and utilize situation as shown in table 1 to carbon and nitrogen sources.
Table 1
NK3-4 is inhibited to pathogenic Fusarium oxysporum for soil of the present invention series bacillus (Paenibacillus terrae), can utilize this characteristic to prevent and treat crop pest, reduces chemical pesticide consumption, alleviates the pollution of chemical pesticide to environment and agricultural-food.
Soil of the present invention series bacillus (Paenibacillus terrae) NK3-4 has the nitrogen in fixed air, decomposes the function of mineral phosphorus (inorganic phosphorus), mineral K and insoluble silicon.
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.
Fig. 3 is the dull and stereotyped antagonism figure of soil series bacillus (Paenibacillus terrae) NK3-4 bacterial strain and pathogen of soybean root rot Fusarium oxysporum.
Embodiment
Technical solution of the present invention is not limited to following cited embodiment, also comprises the arbitrary combination between each embodiment.
Embodiment one: present embodiment soil series bacillus (Paenibacillus terrae) NK3-4, be deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preserving number is CGMCC No.7011.
Present embodiment soil series bacillus (Paenibacillus terrae) NK3-4 sampling position is experiment field (46 ° of 46 ' 02.8 " N in Heilongjiang Province, Kiamusze City, Heilongjiang Province of China institute of land-reclaimable academy of sciences; 130 ° 25 ' 44.5 " E).
Soil series bacillus (Paenibacillus terrae) NK3-4 bacterial strain gathers, Isolation and screening:
In July, 2005, in the continuous cropping Soybean Field of 4 years, select the soybean plant strain of robust growth, uproop, collect root week soil, from the collected specimens of sampling position, each sample is classified respectively, numbered, pack in the sealing polyethylene bag of sterilizing, be placed in and in ice chest, take back laboratory, 4 ℃ of preservations, carry out enrichment incubation in 2 days.By above-mentioned pedotheque 1g, (selectivity potassium felspar sand liquid nutrient medium is by 10.0g sucrose, 0.2g K to put into selectivity potassium felspar sand liquid nutrient medium again
2hPO
4, 0.2gMgSO
47H
2o, 0.2g NaCl, 5.0g CaCO
3, 1.0g feldspar in powder, 0.1g CaSO
4, 2.0g yeast powder and 1L distilled water composition, pH value 7.2) carry out (30 ℃ of enrichment incubations, 200r/min, cultivate 24h), after enrichment incubation, by nutrient solution gradient dilution, each gradient concentration takes out 100 μ L and is inoculated in 3 repetitions of the each gradient of selectivity potassium felspar sand solid medium (agar plate) with coating method, after 30 ℃ of cultivation 48h, select single bacterium colony to carry out purifying, microscope is seen the unicity of looking into bacterium colony, to impure being further purified, until obtain pure bacterial strain.
The Main Pathogenic Bacteria Fusarium oxysporum of the pure bacterial strain of above-mentioned acquisition and pathogen of soybean root rot is carried out to bacteriostatic experiment, therefrom filter out strain excellent Fusarium oxysporum to good antagonistic action.This Fusarium oxysporum is provided by the land-reclaimable academy of sciences in Heilongjiang Province Plant Protection Institute.
Adopt plate face-off culture method.Fusarium oxysporum point is connected on kind of PDA substratum, cultivate after 5d for 25 ℃, first get pathogenic Fusarium oxysporum lawn uniformity with the aseptic cake device of beating, diameter is the bacterium cake of 10mm, bacterium cake is placed in to PDA culture medium culturing ware, plate diameter 9cm, bacterium cake is placed on 3 summits of equilateral triangle centered by the plate center of circle, pathogenic Fusarium oxysporum Jun Bing center and the culture dish center of circle are at a distance of 30mm, get again to be coated with and be inoculated on LA substratum, cultivate 2d for 30 ℃, lawn thickness is consistent, diameter is the PDA substratum plate center that the bacterium cake of the strains tested to be measured of 10mm is placed in the pathogenic Fusarium oxysporum bacterium of above-mentioned inoculation cake, strains tested to be measured and pathogenic Fusarium oxysporum form face-off and cultivate situation.Establish altogether 5 repetitions, after 28 ℃ of cultivation 36h, measure inhibition zone size, select with pathogenic Fusarium oxysporum face-off and cultivate the bacterial strain of inhibition zone maximum, according to 16S rDNA and physiological and biochemical test qualification result, be judged to be the new bacterial strain of a strain in soil class bacillus kind, called after soil series bacillus (Paenibacillus terrae) NK3-4.Soil series bacillus (Paenibacillusterrae) NK3-4 reaches 8.0mm(as shown in Figure 3 to the antibacterial bandwidth of pathogen of soybean root rot Fusarium oxysporum), inhibition is far beyond other strains tested to be measured.
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 Rock Phosphate (72Min BPL) is to grow on the solid plate substratum in unique phosphorus source, containing 1%Ca
3(PO
4)
2phosphorus decomposing nutrient solution in cultivate after 8d, with molybdenum antimony resistance colorimetric method, measuring the concentration of rapid available phosphorus in nutrient solution is 57.6～70.5mg/L.
Soil series bacillus (Paenibacillus terrae) NK3-4 bacterial strain shows the ability of stronger decomposition silicate minerals under shaking flask condition.Take feldspar in powder as substrate, cultivate after 24 hours, the concentration of substratum effective K is 127.3mg/L, thalline effective K concentration is 22.0mg/L, nutrient solution effective K concentration does not more add potassium felspar sand and processes the high 15.0mg/L of contrast, and fixing potassium does not more add the high 4.8mg/L of processing control group of potassium felspar sand in soil series bacillus (Paenibacillus terrae) NK3-4 thalline.
Soil series bacillus (Paenibacillus terrae) NK3-4 bacterial strain is when destroying silicate minerals, also discharge element silicon, take potassium felspar sand as substrate, the concentration that connects the silicon in bacterium processing filtrate is 193.0～226.2mg/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 nutrient solution, and in bacterium liquid, quick-acting nitrogen contents can reach 8.1～11.6mg/L, higher by 368.1% than the control group of inoculation inactivated bacteria processing.
Present embodiment soil series bacillus (Paenibacillus terrae) NK3-4 is reducing applying quantity of chemical fertilizer in the situation that, 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.
Embodiment 1
Picking soil series bacillus (Paenibacillus terrae) NK3-4 lawn is inoculated in potassium felspar sand liquid nutrient medium, and (potassium felspar sand liquid nutrient medium is by 10.0g 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 yeast powder and 1L distilled water composition, pH value 7.2), (control group substratum 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 substratum of K-feldspar powder
2hPO
4, 0.2g MgSO
47H
2o, 0.2g NaCl, 5.0g CaCO
3, 0.1g CaSO
4, 2.0g yeast powder and 1L distilled water composition, pH value is 7.2), with the potassium felspar sand 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 photometry, survey the concentration of potassium in substratum, 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 substratum 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, concentration at the potassium felspar sand substratum effective K of inoculating soil series bacillus (Paenibacillus terrae) NK3-4 is 127.3mg/L, thalline effective K concentration is 22.0mg/L, and the concentration of the control group substratum 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 felspar sand culture medium culturing, 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(culture condition, 200r/min at A Xubei (Ashby) in without nitrogen nutrient solution), 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 nutrient solution to (every liter of phosphorus decomposing nutrient solution contains glucose 10.0g, NaCl0.3g, MnSO
44H
2o0.03g, (NH
4)
2sO
40.5g, KCl0.3g, 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 nutrient solution with molybdenum blue colorimetric method, in the substratum 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% chlorine bleach liquor sterilizes 3 minutes that selected soybean 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 (Paenibacillus terrae) NK3-4 liquid bio preparation (all unitedly calling below NK3-4 liquid bio preparation) 10ml, and to add 10ml sterilized water as a control group, repeat for 9 times, within the 3rd day, survey germinating energy, within the 6th day, survey percentage of germination.
Experimental result is as shown in table 2, and NK3-4 liquid bio preparation can make soybean seeds percentage of germination improve 4.4 percentage points.
Experiment conclusion: NK3-4 liquid bio preparation can do soybean seeds pre-sowing treatment.
Table 2
Embodiment 3
For examination soil, be continuous cropping soil (5 years Soybean Fields of continuous cropping) and main crop soil (Soybean Field that front stubble is 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 soybean seeds sowing by seed coat agent (effective ingredient is 15% derosal, 10% thiram and 10% carbofuran) by the weight ratio Dressing of seed coat agent and soybean seeds 1 ︰ 100; The 2nd group: before soybean seeds sowing, use NK3-4 liquid bio preparation in the ratio Dressing of 1000 grams of soybean seeds of 10ml NK3-4 liquid bio Ji ︰ processed; The 3rd group: using clear water Dressing in proportion 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, all can reduce dry matter accumulation of soybean amount and disease index in main crop soil by NK3-4 liquid bio preparation and seed coat agent, 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, Repone K 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
Within 2012, in experiment field, land reclamation and cultivation academy of sciences rice research institute garden, Heilongjiang Province, carry out soil organism 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 ordinary method, samples, species test during rice harves, in each community, chooses 3m
2harvesting is surveyed and is produced.Species test and surveying produces that result is as shown in table 5, and what conventional fertilizer application enriched siliceous fertilizer and used NK3-4 liquid bio preparation of the present invention processing can obviously improve rice yield constituent element, and paddy rice mean 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% derosal, 10% thiram, 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%), Repone K (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 type is black earth, soybean continuous cropping 6 years, organic content 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 (Paenibacillus terrae) NK3-4 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 Repone K 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 trial plot, 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.Soil moisture deficiency during sowing, 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.Broadcast and with acetochlor 1500ml+, match gram 400g before rear seedling and 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, as seed-fertilizer applying effect of increasing production the best; 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% does seed manure, output is executed and is improved 3.9%～8.2% than routine.
Table 8
Soil series bacillus in above-described embodiment (Paenibacillus terrae) NK3-4 liquid bio preparation adopts following steps preparation:
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 size, 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 ℃, stirring velocity, is that 220r/min and sterile 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, 0.2g MgSO
4, 0.2g NaCl, 5.0g CaCO
3, 1.0g CaSO
4, 2.0g yeast powder, 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 size, be inoculated in the production tank that fermention medium is housed, the volume of producing fermention medium in tank is to produce 70% of tank volume, in fermention medium per ton, contain 1.2kg sucrose, 2.6kg W-Gum, 1.2kg peptone, 0.84kg dry yeast, 0.24kg magnesium sulfate, 0.6kg CaCO
3, 1.2kg K
2hPO
4, 6.3kg soybean cake powder, 18.0kg Semen Maydis powder, 0.9kg yeast, 0.15L defoamer, 1.35kg soya-bean oil, pH value 7.2～7.5 after sterilizing; Then at 28～30 ℃, stirring velocity, be that 250r/min and sterile air intake are that in being cultured to soil series bacillus (Paenibacillus terrae) NK3-4 fermented liquid under the condition of 1 ︰ 0.8,95% above thalline produces brood cell, and brood cell's quantity exceedes 1,000,000,000/ml, obtain soil series bacillus (Paenibacillus terrae) NK3-4 liquid bio preparation.
Soil series bacillus in above-described embodiment (Paenibacillus terrae) NK3-4 solid biologic preparation adopts following steps preparation: soil series bacillus (Paenibacillus terrae) NK3-4 liquid bio preparation is mixed with the mass ratio of the peat composed of rotten mosses 1 ︰ 10 by liquid preparation, and adding the white clay of solid biologic preparation total mass 30%～35% and 10%～15% humic acid as auxiliary material, extruder grain obtains soil series bacillus (Paenibacillus terrae) NK3-4 solid biologic preparation.
Claims (1)
1. a strain soil series bacillus, it is soil series bacillus (Paenibacillus terrae) NK3-4, is deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preserving number is CGMCC No.7011.
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