CN103146622A - Paenibacillus terrae strain - Google Patents

Paenibacillus terrae strain Download PDF

Info

Publication number
CN103146622A
CN103146622A CN2013100974502A CN201310097450A CN103146622A CN 103146622 A CN103146622 A CN 103146622A CN 2013100974502 A CN2013100974502 A CN 2013100974502A CN 201310097450 A CN201310097450 A CN 201310097450A CN 103146622 A CN103146622 A CN 103146622A
Authority
CN
China
Prior art keywords
soil
paenibacillus terrae
series bacillus
paenibacillus
terrae
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CN2013100974502A
Other languages
Chinese (zh)
Other versions
CN103146622B (en
Inventor
于文清
刘文志
李鹏
胡广民
隋文志
赵晓锋
田艳洪
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Heilongjiang Academy of Land Reclamation Sciences
Original Assignee
Heilongjiang Agribusiness Sanlong Biotechnology Co ltd
Utilization Of Agricultural And Livestock Research Institute Heilongjiang Academy Of Land Reclamation Sciences
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Heilongjiang Agribusiness Sanlong Biotechnology Co ltd, Utilization Of Agricultural And Livestock Research Institute Heilongjiang Academy Of Land Reclamation Sciences filed Critical Heilongjiang Agribusiness Sanlong Biotechnology Co ltd
Priority to CN201310097450.2A priority Critical patent/CN103146622B/en
Publication of CN103146622A publication Critical patent/CN103146622A/en
Application granted granted Critical
Publication of CN103146622B publication Critical patent/CN103146622B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Landscapes

  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)
  • Fertilizers (AREA)

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

One strain soil series bacillus
Technical field
The present invention relates to a kind of series bacillus.
Background technology
Class Bacillus (Paenibacillus Ash, Priest﹠amp; Collin, 1994) belong to Firmicutes, bacillus guiding principle, bacillus order, class Bacillaceae.It is shaft-like that cell is, and Gram-positive, feminine gender or variable are moved with peritrichous.Oval gemma is arranged in cyst expanding, on nutrient agar medium without soluble pigment.Amphimicrobian or strictly aerobic.At least, recorded at present 39 kinds, the class bacillus is widely distributed at occurring in nature.
The research of known class bacillus is found that 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 the bacterium thalline again, and can induce host plant to produce disease resistance, can also provide nutrition for host plant by fixed nitrogen and dissolved organic phosphorus, 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 stronger tolerance.And found also 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 the exocellular polysaccharide function in the straw compost of olive squeezing waste material composting; Daane equals to find can the degrade novel species P.naphthalenovorans of naphthalene of a strain at the sabkha plant rhizosphere in 2002; Yoon etc. found that two strains can produce novel species P.kribbensis and the P.terrae of the flocculation agent of efficient flocculating marine alga in 2003; Horn etc. found that a strain can produce N in 2005 in the 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 the HPL-003 bacterial strain can produce zytase, and the 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, because crop is low to its utilization ratio, easy loss, 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﹠amp; 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 the brood cell, when forming the brood cell, the thalline near-end expands, and cyst obviously expands, the 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 formed the circular bacterium colony that diameter is 1~2mm left and right in 5~7 days, single bacterium colony median rise, the 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, gluing the lawn amount more, be cultured to the 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, does not grow in 2%NaCl; And Paenibacillus terrae AM141 gramstaining is variable, and thalline length is 1.3~1.8 μ m * 4~7 μ m, can utilize N.F,USP MANNITOL, grows in 2%NaCl.Therefore, judge that soil series bacillus (Paenibacillus terrae) NK3-4 is a new bacterial strain of strain of soil series bacillus kind.
Soil of the present invention series bacillus (Paenibacillus terrae) NK3-4 gramstaining is positive, and indole reaction is negative, and the M.R test is negative, and the 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 the energy hydrolyzed starch does not utilize N.F,USP MANNITOL; Produce sour aerogenesis when utilizing dextrose plus saccharose, produce not aerogenesis of acid when utilizing lactose, Xylo-Mucine, humic acid, after general the cultivation, the 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 grows in 1%NaCl, does not grow in 2%NaCl; Optimum growth temperature is 30~32 ℃, and the 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 the chemical pesticide consumption, alleviates chemical pesticide to the pollution of 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﹠amp; Collin, 1994), be deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC), the preservation address is the Datun Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, preservation date is on December 17th, 2012, preserving number is CGMCC No.7011.
Description of drawings
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, Heilongjiang Province of China Kiamusze City 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, after the collected specimens of sampling position, each sample is classified respectively, numbered, in the sealing polyethylene bag of the sterilization of packing into, be placed in and take back the laboratory in ice chest, enrichment incubation is carried out in 4 ℃ of preservations in 2 days.With 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 forms, pH value 7.2) carry out (30 ℃ of enrichment incubations, 200r/min, cultivate 24h), after enrichment incubation, with the nutrient solution gradient dilution, each gradient concentration takes out 100 μ L and is inoculated in 3 repetitions of each gradient of selectivity potassium felspar sand solid medium (agar plate) with coating method, select single bacterium colony to carry out purifying after 30 ℃ of cultivation 48h, microscope is seen the unicity of looking into bacterium colony, to impure being further purified, until obtain pure bacterial strain.
The pure bacterial strain of above-mentioned acquisition and the Main Pathogenic Bacteria Fusarium oxysporum of pathogen of soybean root rot are carried out bacteriostatic experiment, therefrom filter out the strain excellent that Fusarium oxysporum is had 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, after 25 ℃ of cultivation 5d, first get pathogenic Fusarium oxysporum lawn uniformity with the aseptic cake device of beating, diameter is the bacterium cake of 10mm, the bacterium cake is placed in PDA culture medium culturing ware, plate diameter 9cm, the bacterium cake is placed on equilateral triangle 3 summits centered by the plate center of circle, pathogenic Fusarium oxysporum bacterium cake center and the culture dish center of circle are at a distance of 30mm, get again to be coated with and be inoculated on the LA substratum, cultivate 2d for 30 ℃, the 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, measure the inhibition zone size after 28 ℃ of cultivation 36h, select with pathogenic Fusarium oxysporum face-off and cultivate the bacterial strain of inhibition zone maximum, be judged to be a new bacterial strain of strain in soil class bacillus kind, called after soil series bacillus (Paenibacillus terrae) NK3-4 according to 16S rDNA and physiological and biochemical test qualification result.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 other strains tested to be measured head and shoulders above.
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, is containing 1%Ca 3(PO 4) 2The phosphorus decomposing nutrient solution in cultivate 8d after, the concentration of measuring rapid available phosphorus in nutrient solution 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 minerals under the 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 in soil series bacillus (Paenibacillus terrae) NK3-4 thalline, fixing potassium does not more add the high 4.8mg/L of processing control group of potassium felspar sand.
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 the 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 the nitrogen nutrient solution, and in bacterium liquid, quick-acting nitrogen contents can reach 8.1~11.6mg/L, and the control group of processing than the inoculation inactivated bacteria is high by 368.1%.
Series bacillus (Paenibacillus terrae) NK3-4 in present embodiment soil still can make paddy rice, corn, Soybean and Other Crops volume increase in the situation that reduce applying quantity of chemical fertilizer, and particularly to be used in conjunction with the increasing production of rice amplitude more remarkable with siliceous fertilizer.
Embodiment 1
Picking soil series bacillus (Paenibacillus terrae) NK3-4 lawn is inoculated in the potassium felspar sand liquid nutrient medium, and (the 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 forms, pH value 7.2), (the control group substratum is by 10g sucrose, 0.2g K in addition soil series bacillus (Paenibacillus terrae) NK3-4 to be inoculated in not 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 forms, pH value is 7.2),, test and establish 4 repetitions as blank with the potassium felspar sand liquid nutrient medium that do not connect bacterium, cultivate 24h under 30 ℃, 200r/min condition, standby.
The bacterium liquid of getting above-mentioned cultivation 24h dilutes 10 times, survey the concentration of potassium in substratum with flame photometry, 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 the 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, and 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, measure the concentration of quick-acting silicon in filtrate with molybdenum blue spectrophotometric method, result shows that the concentration of silicon in the filtrate of inoculating 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, and is 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 the 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 the phosphorus decomposing nutrient solution (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, cultivate 8d under 30 ℃, 200r/min condition after, 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 the bacterium control group and to be 13.3mg/L, the former is 4.8 times of the latter.
Embodiment 2
It is 0.1% chlorine bleach liquor sterilization 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 sterilization circle filter paper, 15/plate, add soil series bacillus (Paenibacillus terrae) NK3-4 liquid bio preparation (the following NK3-4 liquid bio preparation that all unitedly calls) 10ml, and to add the 10ml sterilized water as a control group, 9 repetitions, surveyed germinating energy on the 3rd day, the 6th day survey percentage of germination.
Experimental result is as shown in table 2, and NK3-4 liquid bio preparation can make the soybean seeds percentage of germination improve 4.4 percentage points.
Experiment conclusion: NK3-4 liquid bio preparation can be done the soybean seeds pre-sowing treatment.
Table 2
Figure BDA00002962466300061
Embodiment 3
Be continuous cropping soil (5 years Soybean Fields of continuous cropping) and main crop soil (front stubble is the Soybean Field of corn) for examination soil, front stubble is not for the sickle-like bacteria spore being detected in the soyabean field of corn.Get continuous cropping soil and sieve with main crop folk song is dried, be loaded in 2.2L basin alms bowl, every alms bowl fills native 2.35kg.Experiment is divided into 3 groups, the 1st group: the weight ratio Dressing of pressing kind of clothing agent and soybean seeds 1 ︰ 100 before the soybean seeds sowing with kind of clothing agent (effective ingredient is 15% derosal, 10% thiram and 10% carbofuran); The 2nd group: use NK3-4 liquid bio preparation in the ratio Dressing of 10ml NK3-4 liquid bio Ji ︰ 1000 gram soybean seeds processed before the soybean seeds sowing; The 3rd group: with clear water Dressing in proportion as the blank group.6 seeds of every alms bowl sowing, the rear field planting of emerging, 4 repetitions are established in every alms bowl 2 strains.Measure plant height, dry-matter accumulation amount, dross situation, root rot disease index period in soybean the 3rd compound leaf and the 5th compound leaf.
Experimental result is as shown in table 3, all can reduce dry matter accumulation of soybean amount and disease index in main crop soil with NK3-4 liquid bio preparation and the agent of kind clothing, 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 plant the clothing agent to beans seedling dry-matter accumulation amount, root nodule dry weight and quantity all higher than the blank group, and disease index is all lower than the blank group.The result of use of NK3-4 liquid bio preparation slightly is worse than kind of a clothing agent 3 compound leaf phases, is better than kind of a clothing agent 5 compound leaf phase effects.Experiment shows the effect lasting stability of using NK3-4 liquid bio preparation.
Table 3
Figure BDA00002962466300071
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.3 repetitions are established in the residential quarter experiment, and random district group is arranged, and the residential quarter area is 16.25m 2Conventional fertilizer application: urea 40kg/hm 2, diammonium phosphate 150kg/hm 2, Repone K 40kg/hm 2Conventional fertilizer application is also dressed seed with NK3-4 liquid bio preparation, and NK3-4 liquid bio preparation consumption is the 10ml/kg seed.
With NK3-4 liquid bio preparation Dressing, the 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
Carried 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 in 2012 in experiment field, Heilongjiang Province land reclamation and cultivation rice research institute of academy of sciences garden.Residential quarter area 24m 2, line-spacing 25cm, the cave is apart from 10cm.
Experiment arranges 9 experimental group, 3 repetitions, and for cultivating rice 10, each experimental group fertilising situation is as shown in table 4 for the rice varieties that tries the water.
Table 4
Figure BDA00002962466300081
Field management measure is all undertaken by locality plantation ordinary method, takes a sample during rice harves, species test, chooses 3m in each residential quarter 2Harvesting is surveyed and is produced.What species test and survey to produce result as shown in table 5, conventional fertilizer application enriched siliceous fertilizer and used that NK3-4 liquid bio preparation of the present invention processes can obviously improve the rice yield constituent element, and the 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
Figure BDA00002962466300082
Embodiment 6
Be 15% derosal, 10% thiram, 10% carbofuran for planting experimentally clothing agent effective ingredient; (executing can rich board: N13%, P for commercial anti continuous and alternative-year composite fertilizer special 2O 525%, K 2O10%, little 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.
The 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, 4 repetitions, residential quarter area 16.25m 2, soil series bacillus (Paenibacillus terrae) NK3-4 solid biologic preparation (the following NK3-4 solid biologic preparation that all unitedly calls) per hectare consumption is 45kg, and applies so that the base manure form is disposable after the chemical fertilizer mixing.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% kind clothing 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 the individual plant pod is heavy, stem is heavy also in the prostatitis.Experimental group 1 output is the highest, than conventional fertilizer application (experimental group 4) volume increase 10.06%, than kind of a clothing 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
Figure BDA00002962466300091
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.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.
Employing residential quarter experiment, 6 row districts, the long 8m of row repeats for 3 times, the group arrangement of random district, residential quarter area 31.2m 2Experiment is established 10 groups, and the fertilising situation is as shown in table 7.
Table 7
Figure BDA00002962466300101
Sowing on May 7, artificial ditching-fertilizing, equidistant program request, density 60,000 strains/hm 2Carry out enclosed weeding, 4~5 leaf phase of corn subsoiling with acetochlor 1500ml+ match gram 400g before broadcasting rear seedling.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% than the conventional fertilizer application contrast, NK3-4 solid biologic preparation is made seed manure, leaf fertilizer, a significant difference that topdresses, and is best with 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 than conventional fertilizer application contrast volume increase 10%.Normally use in the situation that topdress, calculate by corn conventional fertilizer application amount: soil series bacillus biosolids preparation 25%+ chemical fertilizer 75% is done seed manure, and output is executed than routine and improved 3.9%~8.2%.
Table 8
Figure BDA00002962466300102
Soil series bacillus in above-described embodiment (Paenibacillus terrae) NK3-4 liquid bio preparation adopts the following steps preparation:
A, soil series bacillus (Paenibacillus terrae) NK3-4 is seeded to liquid nutrient medium, cultivate 13h under the 180r/min condition, then be inoculated in the seeding tank that liquid nutrient medium is housed by 0.5% inoculum size, in seeding tank, the volume of liquid nutrient medium is 70% of seeding tank volume, is that 220r/min and sterile air intake are to cultivate 11h under the condition of 1 ︰ 0.8 at 28~30 ℃, stirring velocity; 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, be inoculated in the production tank that fermention medium is housed by 5% inoculum size through the soil of seed tank culture series bacillus (Paenibacillus terrae) NK3-4, produce the volume of fermention medium in tank for producing 70% of tank volume, contain 1.2kg sucrose in fermention medium per ton, 2.6kg W-Gum, 1.2kg peptone, 0.84kg dry yeast, 0.24kg sal epsom, 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 sterilization; Then 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 the brood cell at 28~30 ℃, stirring velocity, and brood cell's quantity surpasses 1,000,000,000/ml, namely obtains 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 the following steps preparation: soil series bacillus (Paenibacillus terrae) NK3-4 liquid bio preparation is mixed by the mass ratio of liquid preparation with the peat composed of rotten mosses 1 ︰ 10, 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.
Figure IDA00002962467100021

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.
CN201310097450.2A 2013-03-25 2013-03-25 Paenibacillus terrae strain Active CN103146622B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201310097450.2A CN103146622B (en) 2013-03-25 2013-03-25 Paenibacillus terrae strain

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201310097450.2A CN103146622B (en) 2013-03-25 2013-03-25 Paenibacillus terrae strain

Publications (2)

Publication Number Publication Date
CN103146622A true CN103146622A (en) 2013-06-12
CN103146622B CN103146622B (en) 2014-05-07

Family

ID=48544945

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201310097450.2A Active CN103146622B (en) 2013-03-25 2013-03-25 Paenibacillus terrae strain

Country Status (1)

Country Link
CN (1) CN103146622B (en)

Cited By (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104988101A (en) * 2015-08-03 2015-10-21 黑龙江省科学院微生物研究所 Bacillus cereus and application of bacillus cereus as agricultural biological agent
CN105565936A (en) * 2016-02-27 2016-05-11 黑龙江省农垦科学院农畜产品综合利用研究所 Special bulk blended fertilizer added with biological preparation for nano rice
CN106962115A (en) * 2017-04-05 2017-07-21 周兴虎 A kind of method for improving willow resistance against diseases
CN108034609A (en) * 2017-12-29 2018-05-15 中国农业大学 One plant of series bacillus bacterial strain T1-1 with phosphate solubilization and its application
CN109112087A (en) * 2018-08-21 2019-01-01 甘肃省农业科学院植物保护研究所 One plant of soil series bacillus YC16-08 and its application
CN110627879A (en) * 2019-10-22 2019-12-31 黑龙江省农垦科学院 Plant disease-resistant protein Ati, coding gene and application
CN111484948A (en) * 2019-01-28 2020-08-04 福建省农业科学院农业生物资源研究所 Root-promoting phosphate-solubilizing nitrogen-fixing bacillus and application thereof
CN111484952A (en) * 2019-01-28 2020-08-04 福建省农业科学院农业生物资源研究所 Bacillus for dissolving phosphorus and fixing nitrogen and application thereof
CN111954712A (en) * 2018-02-07 2020-11-17 瓦拉格罗股份公司 Novel Paenibacillus polymyxa and use thereof
CN114605203A (en) * 2022-04-02 2022-06-10 湖南云谷生物科技发展有限公司 Compound microbial fertilizer and application thereof
CN115044497A (en) * 2022-05-16 2022-09-13 漳州三炬生物技术有限公司 Multifunctional Paenibacillus anggi, composite microbial agent and application thereof
CN115725452A (en) * 2022-09-21 2023-03-03 山东佰渥康生物科技有限公司 Paenibacillus atrophaeus, microbial inoculum and application thereof
CN117467557A (en) * 2023-06-13 2024-01-30 深圳华大生命科学研究院 Paenibacillus strain with growth promoting and biocontrol functions and application thereof

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102851250A (en) * 2012-09-18 2013-01-02 山东农业大学 Rhizosphere antagonistic bacterium paenibacillus polymyxa YC0136 and application thereof

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102851250A (en) * 2012-09-18 2013-01-02 山东农业大学 Rhizosphere antagonistic bacterium paenibacillus polymyxa YC0136 and application thereof

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
JUNG-HOON YOON 等: "Paenibacillus kribbensis sp. nov. and Paenibacillus terrae sp. nov., bioflocculants for efficient harvesting of algal cells", 《INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY》 *
SANG HEUM SHIN 等: "Genome Sequence of Paenibacillus terrae HPL-003, a Xylanase-Producing Bacterium Isolated from Soil Found in Forest Residue", 《JOURNAL OF BACTERIOLOGY》 *
YU,W.Q.等: "JX914567.1", 《GENBANK》 *

Cited By (17)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104988101B (en) * 2015-08-03 2018-03-06 黑龙江省科学院微生物研究所 One plant of bacillus cereus and its purposes as agro-ecology preparation
CN104988101A (en) * 2015-08-03 2015-10-21 黑龙江省科学院微生物研究所 Bacillus cereus and application of bacillus cereus as agricultural biological agent
CN105565936A (en) * 2016-02-27 2016-05-11 黑龙江省农垦科学院农畜产品综合利用研究所 Special bulk blended fertilizer added with biological preparation for nano rice
CN106962115A (en) * 2017-04-05 2017-07-21 周兴虎 A kind of method for improving willow resistance against diseases
CN108034609A (en) * 2017-12-29 2018-05-15 中国农业大学 One plant of series bacillus bacterial strain T1-1 with phosphate solubilization and its application
CN111954712A (en) * 2018-02-07 2020-11-17 瓦拉格罗股份公司 Novel Paenibacillus polymyxa and use thereof
CN111954712B (en) * 2018-02-07 2024-01-02 瓦拉格罗股份公司 Novel Paenibacillus polymyxa and application thereof
CN109112087A (en) * 2018-08-21 2019-01-01 甘肃省农业科学院植物保护研究所 One plant of soil series bacillus YC16-08 and its application
CN109112087B (en) * 2018-08-21 2021-06-29 甘肃省农业科学院植物保护研究所 Paenibacillus terrae YC16-08 and application thereof
CN111484952A (en) * 2019-01-28 2020-08-04 福建省农业科学院农业生物资源研究所 Bacillus for dissolving phosphorus and fixing nitrogen and application thereof
CN111484948A (en) * 2019-01-28 2020-08-04 福建省农业科学院农业生物资源研究所 Root-promoting phosphate-solubilizing nitrogen-fixing bacillus and application thereof
CN110627879A (en) * 2019-10-22 2019-12-31 黑龙江省农垦科学院 Plant disease-resistant protein Ati, coding gene and application
CN114605203A (en) * 2022-04-02 2022-06-10 湖南云谷生物科技发展有限公司 Compound microbial fertilizer and application thereof
CN115044497A (en) * 2022-05-16 2022-09-13 漳州三炬生物技术有限公司 Multifunctional Paenibacillus anggi, composite microbial agent and application thereof
CN115044497B (en) * 2022-05-16 2023-03-10 漳州三炬生物技术有限公司 Multifunctional Paenibacillus anggi, composite microbial agent and application thereof
CN115725452A (en) * 2022-09-21 2023-03-03 山东佰渥康生物科技有限公司 Paenibacillus atrophaeus, microbial inoculum and application thereof
CN117467557A (en) * 2023-06-13 2024-01-30 深圳华大生命科学研究院 Paenibacillus strain with growth promoting and biocontrol functions and application thereof

Also Published As

Publication number Publication date
CN103146622B (en) 2014-05-07

Similar Documents

Publication Publication Date Title
CN103146622B (en) Paenibacillus terrae strain
US8518428B2 (en) Antagonistic bacteria for controlling the Fusarium wilt of continuous cropping banana and their microbial organic fertilizer
CN106591193B (en) One plant of bacillus amyloliquefaciens with the degeneration-resistant effect of wide spectrum growth-promoting
CN103964946B (en) A kind of paddy rice Special compound microbial fertilizer and preparation method thereof
CN104928201B (en) A kind of bacillus amyloliquefaciens HN-11 and its microbial inoculum
CN101781569B (en) Soil improvement and anti continuous cropping multifunctional biological agent
CN107502572B (en) Application of bacillus licheniformis in straw degradation, microbial agent containing bacillus licheniformis and application of bacillus licheniformis
CN101220335B (en) Bacterial strain for preventing and controlling tobacco black shank and bacterium agent thereof
CN104928212A (en) Bacillus megaterium strain X3 and preparation method and application thereof
CN103184184B (en) Streptomyces albidoflavus and applications thereof
CN103194405B (en) Growth-promoting bacteria for promoting ginger growth and preventing and controlling continuous cropping ginger soil-borne wilt and microorganism organic fertilizer produced from growth-promoting bacteria
CN104164394A (en) Antagonistic phytopathogen strain and application thereof
CN103141517B (en) Paenibacillus terrae biological agent and application thereof in agriculture
CN102787090B (en) Alfalfa root growth-promoting rhizobacteria MJM-5 and application of alfalfa root growth-promoting rhizobacteria MJM-5
CN101948780B (en) Antagonist bacterium for preventing and treating continuous cropping hot pepper epidemic disease and microbial organic fertilizer thereof
CN101497541B (en) Efficient disease-resistant phosphate solubilizing bacterial manure for tobacco and production method thereof
CN103468620A (en) Streptomyces albidoflavus strain and application thereof to control cucumber downy mildew
CN106305793A (en) Compound bacterial manure used for prevention and control of cotton verticillium wilt and the preparation method thereof
CN101886055B (en) Antagonistic bacteria NJL-14 for preventing and controlling continuous-cropping tobacco bacterial wilt
CN106635903A (en) Growth-promoting bacteria combination for enhancing salt tolerance of crops in moderate-severe saline and alkaline lands
CN102399713A (en) Bacillus subtilis HL-1 and application thereof in respect of soil phosphate dissolving
CN103952355A (en) Compound microbial phosphate-solubilizing bacterial agent and preparation method thereof
CN102827793A (en) Medicago pseudomonas strain capable of producing ACC (1-aminocyclopropane-1-carboxylate) deaminase and application of medicago pseudomonas strain
CN103468591B (en) Salt-tolerant trichoderma pleuroticola strain and application thereof
CN102174428A (en) Antagonistic bacteria for preventing bacterial wilt of potatoes and microorganism organic fertilizer thereof

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
C14 Grant of patent or utility model
GR01 Patent grant
TR01 Transfer of patent right
TR01 Transfer of patent right

Effective date of registration: 20240326

Address after: No.101, Xiangfu Road, Xiangfang District, Harbin City, Heilongjiang Province

Patentee after: HEILONGJIANG ACADEMY OF LAND RECLAMATION SCIENCES

Country or region after: China

Address before: 154007 No. 382, Anqing street, Jiamusi, Heilongjiang

Patentee before: UTILIZATION OF AGRICULTURAL AND LIVESTOCK Research Institute HEILONGJIANG ACADEMY OF LAND RECLAMATION SCIENCES

Country or region before: China

Patentee before: HEILONGJIANG AGRIBUSINESS SANLONG BIOTECHNOLOGY Co.,Ltd.