CN1276074C - Lichen bacillus for the production of phytocytomine and plant growth regulator - Google Patents
Lichen bacillus for the production of phytocytomine and plant growth regulator Download PDFInfo
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- CN1276074C CN1276074C CN 200310114330 CN200310114330A CN1276074C CN 1276074 C CN1276074 C CN 1276074C CN 200310114330 CN200310114330 CN 200310114330 CN 200310114330 A CN200310114330 A CN 200310114330A CN 1276074 C CN1276074 C CN 1276074C
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- zeatin
- bacillus licheniformis
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- 238000004519 manufacturing process Methods 0.000 title claims abstract description 24
- 239000005648 plant growth regulator Substances 0.000 title claims description 34
- 241000193830 Bacillus <bacterium> Species 0.000 title description 4
- UZKQTCBAMSWPJD-UQCOIBPSSA-N trans-Zeatin Natural products OCC(/C)=C\CNC1=NC=NC2=C1N=CN2 UZKQTCBAMSWPJD-UQCOIBPSSA-N 0.000 claims abstract description 42
- 229940023877 zeatin Drugs 0.000 claims abstract description 42
- UZKQTCBAMSWPJD-FARCUNLSSA-N trans-zeatin Chemical compound OCC(/C)=C/CNC1=NC=NC2=C1N=CN2 UZKQTCBAMSWPJD-FARCUNLSSA-N 0.000 claims abstract description 41
- 241000194108 Bacillus licheniformis Species 0.000 claims abstract description 33
- GOSWTRUMMSCNCW-HNNGNKQASA-N 9-ribosyl-trans-zeatin Chemical compound C1=NC=2C(NC\C=C(CO)/C)=NC=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O GOSWTRUMMSCNCW-HNNGNKQASA-N 0.000 claims abstract description 22
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- 238000000034 method Methods 0.000 claims abstract description 17
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- FAIXYKHYOGVFKA-UHFFFAOYSA-N Kinetin Natural products N=1C=NC=2N=CNC=2C=1N(C)C1=CC=CO1 FAIXYKHYOGVFKA-UHFFFAOYSA-N 0.000 description 1
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- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 1
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- 102000015439 Phospholipases Human genes 0.000 description 1
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- Agricultural Chemicals And Associated Chemicals (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
Zeatin, isopentenyl adenine and zeatin riboside have obvious effect of growth promotion and yield increase for crops. The present invention provides bacillus licheniformis CGMCC No. 1009 for producing zeatin, isopentenyl adenine and zeatin riboside, a plant growth regulating agent containing the bacillus licheniformis and metabolic products thereof, and a method for producing the plant growth regulating agent. The method comprises five steps: (1) production strain cultivation, (2) strain cultivation of a shake flask, (3) fermentation cultivation of a fermentation tank, (4) thin film concentration and (5) emulsification processing. The present invention is characterized in that the bacillus licheniformis is used as a strain source in the step (1); the temperature is from 27 to 37 DEG C, and the pH value is from 7 (+/-) 0.5 in the process of cultivation in steps (1), (2) and (3); the pH value of the concentrated solution obtained from the step (4) is regulated to 5 (+/-) 0.5 in the step (5); an emulsifying agent is added until particles in the concentrated solution are stably suspended. The present invention also provides the plant growth regulating agent for the use of the crops. The plant growth regulating agent has the advantages of simple production technology, low cost, easy use and obvious effect.
Description
Technical field
The present invention relates to microorganism field, more particularly, relate to the plant-growth regulator that is used to produce the Bacillus licheniformis of phytocytomine and contains this bacillus and above-mentioned phytocytomine (zeatin, isopentenyl gland purine and ribosylzeatin).
Background technology
Zeatin, isopentenyl gland purine and ribosylzeatin all belong to the cytokinin growth substance.Phytokinin is one of five big natural phytohormones (being growth hormone, Plant hormones regulators,gibberellins, phytokinin, dormin and ethene) of having known of present people.Its physiological action is: (1) pair cell mitogen and differentiation have obvious effects.(2) inducing cell increases.(3) can remove apical dominance, promote the lateral bud growth.(4) some seed and bud there is the effect that promotes to sprout breaking dormancy.(5) reduce chlorophyllous hydrolysis, postpone leaf senile.Phytokinin can be widely used in cash crop such as field crop, vegetables, fruit, and significantly improves output and improve quality.This class medicament originates in the discovery of the growth hormone thirties in 20th century, and growth hormone ubiquity, content in plant materials is atomic, but it is the necessary for plant growth material.The purpose of research plant hormone, be not only in order to disclose its mechanism in the coordinate plant growth growth course and the regularity of regulating control, the more important thing is the utilisation technology of exploring plant growth substance, make it by producing needs, the regulation and control growth and development of plant improves quality and the output of crop.The application of plant hormone also provides some conveniences for people in plant growing and breeding.In some cases, reasonably the production effect of appliable plant growth substance even ratio cultivation are faster with breeding.
Phytokinin is divided into 15 kinds again, and zeatin, kinetin, isopentenyl gland purine, ribosylzeatin etc. are wherein arranged, and especially the effect with zeatin is best, but price is also the most expensive.The natural corn element of the U.S. extracts from green corn, from more than 60000 kilograms of green corns, can only extract 1 gram zeatin, every gram price is 50,000 yuans, therefore, only plants middle use at bottle, generally only be used for the phytokinin Its Mechanisms, the field can't be used at all.At present, domestic have report to utilize actinomycetes to produce zeatin and isopentenyl gland purine, and pesticide registration registration content is 1ppm, just can only reach lower concentration.And its fermentation period is longer, generally will be more than 4 days.So its cost is also higher relatively.
Summary of the invention
The objective of the invention is to provide a kind of Bacillus licheniformis that is used to produce phytocytomine, contain this Bacillus licheniformis and, and this plant-growth regulator is used to promote the purposes of crop growth and volume increase as the plant-growth regulator of zeatin, isopentenyl gland purine and the ribosylzeatin of its meta-bolites and the method that is used to produce this plant-growth regulator.
The inventor kills in the acarid bacterial strain process in screening, evenly is sprayed on the string bean tetranychid respectively with the different strains fermented liquid.After 10 days, find that wherein strain bean seedlings look very special, lusty strong again, leaf is very big.Subsequently, respectively wheat, corn, grass seeds, cucumber are carried out pot experiment with this bacterial strain fermentation liquor.Test-results shows: fermented liquid and clear water contrast difference highly significant, 40 times of liquid of this fermented liquid have very strong stimulating growth effect to wheat, corn, cucumber.When maize seedling length to the in the time of 18 days, the young plant of spraying fermented liquid exceeds clear water and contrasts nearly one times (8 centimetres), and very sturdy.Fermented liquid has the very strong bud effect that inspires to grass seeds, and percentage of germination can improve about 50%.Thereby obtain this bacterial strain.Be accredited as Bacillus licheniformis (BacillusLicheniformis) through Institute of Microorganism, Academia Sinica.Strain properties: cell is shaft-like, and Gram-positive, size are 0.8 * 1.5～3.5 μ m, produces nearly middle ellipticity gemma of giving birth to, and sporangiocyst expands slightly.No Poly-salt (PHB) particle in the cell.Bacterium colony on bouillon media be flat, the edge is irregular, white, surface irregularity gauffer, colony diameter is 3mm after 24 hours.Produce acetyl methyl carbinol (V-P reacting positive), pH5.4.The catalase positive.The lecithinase positive, reduction nitrate.In 7%NaCl and pH5.7, grow battalion's amphimicrobian life.Produce acid from glucose, wood sugar and N.F,USP MANNITOL, but do not produce acid from pectinose.Litmus milk is reduction and peptonizes reaction, starch-splitting and casein, and consume Citrate trianion.In recent years, warp, obtained a strain at last and produces plain height, produces spore height, the stable superior strain of tiring by separating with ultraviolet mutagenesis, monospore the continuous repeated screening of bacterial strain.This product has enhancing photosynthesis, and the stimulating plant cell fission promotes chlorophyll to form, and quickens plant metabolism and proteinic synthetic, impels the crop early maturing and high yield, increases the sugar degree of melon dish, improves the anti-ageing cold tolerance of plant disease-resistant.Very obvious to the melon-fruit-like vegetable preventing disease production-increasing effect, amount of increase in production is at 20-30%.So the inventor is sent to the common micro-organisms center preservation of Chinese microbial preservation management committee on September 24th, 2003 with this bacterial strain, the specific name of its suggestion is Bacillus licheniformis (BacillusLicheniformis), and it is registered on the books and is numbered CGMCC No.1009.
In addition, the inventor can clearly promote the mechanism of plant-growth to further investigate to this Bacillus licheniformis CGMCC No.1009, found that this Bacillus licheniformis CGMCCNo.1009 can secrete a large amount of zeatin, isopentenyl gland purine and ribosylzeatin.And above-mentioned three kinds of compositions all are effectively plant growth hormoness, and therefore, the fermented liquid of Bacillus licheniformis CGMCCNo.1009 itself is a kind of good plant-growth regulator.
Because above-mentioned result of study, thereby finished the present invention.
Technical scheme of the present invention is as follows:
(1) a kind of Bacillus licheniformis that is used to produce phytocytomine is characterized in that, it is Bacillus licheniformis CGMCC No.1009, and said phytocytomine is zeatin, isopentenyl gland purine and ribosylzeatin.
(2) one plant growth regulators is characterized in that, wherein contain Bacillus licheniformis CGMCC No.1009 and as zeatin, isopentenyl gland purine and the ribosylzeatin of its meta-bolites.
The production method of (3) one plant growth regulators, comprise: 1. produce spawn culture, 2. shake a bottle spawn culture, 3. ferment tank cultivate, 4. thin film concentration, 5. five steps of emulsification, it is characterized in that, use Bacillus licheniformis CGMCC No.1009 as the bacterial classification source in 1. in step; Step 1., 2., the temperature that adopts in 3. the culturing process is 27～37 ℃, the pH value is 7 ± 0.5; The pH value of the concentrated solution that 4. step is obtained in 5. in step is adjusted to 5 ± 0.5, and adds emulsifying agent to the particulate in the concentrated solution and can stably suspend.
As the production method of the described plant-growth regulator of technical scheme (3), it is characterized in that (4) 3. wherein said step is divided into the 3. medium-sized ferment tank cultivation of a again; 3. two steps of b large fermentation tank fermentation culture.
As the production method of the described plant-growth regulator of technical scheme (3), it is characterized in that (5) prescription of the substratum that uses is when 1. production spawn culture of step:
Composition
Weight %
Sucrose 1～3
Peptone 0.1～0.5
Yeast powder 0.3～0.5
Extractum carnis 0.3～0.7
Agar 1～3
(NH
4)
2SO
4 0.3～0.5
K
2HPO
2 0.03～0.05
MgSO
4·7H
2O 0.02～0.05
MnSO
4 0.001～0.003
Water surplus (supplying 100%),
At the step prescription that shakes the substratum that uses when bottle spawn culture and step ferment tank is 3. cultivated 2. be:
Composition
Weight %
Sucrose 2～6
Soybean cake powder 2～5
Cottonseed cake powder 1～3
Peptone 0.1～0.5
Extractum carnis 0.3～0.7
Yeast powder 0.3～0.7
NaCl 0.1～0.6
(NH
4)
2SO
4 0.3～0.7
K
2HPO
2 0.02～0.05
MgSO
4·7H
2O 0.02～0.05
MnSO
4 0.001～0.003
Water surplus (supplying 100%).
As the production method of the described plant-growth regulator of technical scheme (3), it is characterized in that (6) emulsifying agent that adds in 5. in step is 0203B, its consumption is 1～3 weight % of plant-growth regulator.
As the production method of the described plant-growth regulator of technical scheme (3), it is characterized in that (7) 1. step carries out in incubator, cultivated 24～36 hours that the slant strains that obtains is preserved under 0～4 ℃ temperature, in 1 year, use; 2. step carries out on shaking table, and shaking speed is 150～300rpm, finishes when the gemma number that records with microscopy reaches 10～2,000,000,000/ml to cultivate; 3. step carries out in the industrial fermentation jar, the air air flow be 15～100 tank volume/hour, the gemma number that records with microscopy reach 4,000,000,000/more than the ml, the content of zeatin reaches 40 μ g/ml and finishes cultivation when above simultaneously, put jar, and then carry out step 4. with step 5..
(8) the described plant-growth regulator of technical scheme (2) is used to promote the purposes of crop growth and volume increase.
(9) be used to promote the purposes of crop growth and volume increase as the described plant-growth regulator of technical scheme (8), it is characterized in that wherein said farm crop are food crop, oil crops, vegetables, melon and fruit, cotton, flowers, tobacco and fruit tree.
At length explain the present invention below.
Bacillus licheniformis CGMCC No.1009 is that the inventor comes out by the great deal of experimental screening and culturing.This bacillus can secrete zeatin, isopentenyl gland purine and ribosylzeatin specifically in a large number, with the content of zeatin for the highest.
The zeatin chemical name is: 6-(the anti-butenyl of 4-hydroxy-3-methyl-2-) aminopurine, popular name: Zeatin molecular formula: C
10H
13N
5The O molecular structural formula:
The isopentenyl gland purine chemical name is: 6-(γ, γ-dimethyl-allyl) amido purine, popular name: DMAA, IPA, 2iPA molecular formula: C
10H
13N
5, molecular structural formula:
The Bacillus licheniformis CGMCC No.1009 that in the plant-growth regulator described in the technique scheme (2), is contained and do not need to do particular determination as the ratio and the concentration of zeatin, isopentenyl gland purine and the ribosylzeatin of its meta-bolites, because zeatin, isopentenyl gland purine and ribosylzeatin all are plant growth hormoness, no matter its ratio how, all can reach the effect that promotes plant-growth.These several compositions are all secreted out by Bacillus licheniformis CGMCC No.1009, and this bacillus itself does not play remarkable influence to usefulness, so its ratio does not need to limit yet.In addition, the working concentration of zeatin is very low, even if also can obtain significantly to promote growth result with 0.1ppm even lower concentration use, and utilize Bacillus licheniformis CGMCC No.1009 to produce zeatin, isopentenyl gland purine and ribosylzeatin and the mixture that obtained of directly will ferment to use as growth regulator be that the present invention creates, so do not need the concentration of zeatin, isopentenyl gland purine and ribosylzeatin is made qualification.But the concentration of zeatin preferably more than 0.1ppm, so just guarantees to obtain than obvious effects.Because zeatin isopentenyl gland purine and ribosylzeatin all are the meta-bolitess of Bacillus licheniformis, and zeatin has accounted for major part therein, reach just passable more than the 40 μ g/ml so in technical scheme (7), only need record the content of zeatin, do not need to limit respectively the content of isopentenyl gland purine and ribosylzeatin.
In addition, technical scheme (3)～(7) all are the production methods of plant-growth regulator.Wherein, the feature in the technical scheme (3) is the necessary technology feature, and the feature in the other technologies scheme is supplementary features or the further feature that limits.Adopt Bacillus licheniformis CGMCCNo.1009 as the bacterial classification source in technical scheme (3), existence and the breeding under the condition of pH 7 ± 0.5 and 27～37 ℃ that suit of this bacterial classification is so pH value and temperature are prerequisites.And the substratum that uses in culturing process can be known various substratum, as long as enough nutritive substances are arranged.Stock trick art scheme (3) does not need substratum is made qualification, but the inventor finds out most preferred substratum, now it is documented in the technical scheme (5).The incubation time of each culturing step need not done strict the qualification yet, even because the low effect that promotes plant-growth of yet receiving of zeatin content.But the concentration of zeatin preferably more than 0.1ppm, so that obtain more preferably to reach more than the 40 μ g/ml (40ppm) than obvious effects, so more helps storing and transportation, can be made into different concentration in use as required.The pH value of the concentrated solution that 4. step is obtained in 5. in the step of technical scheme (3) is adjusted to pH 5 ± 0.5th, and is necessary, because can suppress the growth of assorted bacterium like this, thereby guarantees that zeatin, isopentenyl gland purine and ribosylzeatin can be steady in a long-term.In addition, the purpose of adding emulsifying agent is to make the nutrient solution as plant-growth regulator can keep uniform suspended state.
Technical scheme (4) is that 3. the step in the technical scheme (3) 3. is divided into the medium-sized fermentor tank of a (for example 500L) fermentation and 3. two steps of the large fermentation tank of b (for example 20000L) fermentation.But when actual production, can also be divided into more fermentation step.
Finish to cultivate when in technical scheme (7), the step concluding time 2. being defined as the gemma number and reaching 10～2,000,000,000/ml, and the concentration of unqualified zeatin, because step product 2. is not directly to provide as conditioning agent to use, it providing enough bacterial classifications for next procedure, as long as it guarantee.And step product 3. is directly to provide as conditioning agent to use, so preferably simultaneously the content of gemma number and zeatin is limited.Under general situation, the incubation time of each step only needed get final product in 24～48 hours.But preferably gemma number or the gemma number by microscopy adds that zeatin concentration limits the concluding time, and be so more reasonable.
In addition, plant-growth regulator of the present invention can use on all plants, and wherein the more obvious plant of usefulness is food crop, oil crops, vegetables, melon and fruit, cotton, flowers, tobacco and fruit tree etc.
Compare with the state of the art, the present invention has following beneficial effect:
1. Bacillus licheniformis CGMCC No.1009 of the present invention can secrete a large amount of zeatin, isopentenyl gland purine and ribosylzeatin specifically, and these several compositions all are effectively plant growth hormoness;
2. have the various plants phytokinin, its usefulness is apparently higher than the zeatin of the chemosynthesis of single component;
3. fermenting process is simple, speed is fast, and its cost is far below the zeatin that extracts from green corn.
4. easy to use, product is stablized, and can deposit more than 2 years.
Embodiment
Enumerate embodiment and application examples below and explain the present invention particularly.
Embodiment 1
In all each culturing steps, the pH=7 of substratum ± 0.1, temperature is 32 ℃.
Step 1.-produce spawn culture
Use Bacillus licheniformis CGMCC No.1009 as the bacterial classification source, the prescription of used substratum is:
Composition
Weight %
Sucrose 2
Peptone 0.3
Yeast powder 0.4
Extractum carnis 0.5
Agar 2
(NH
4)
2SO
4 0.4
K
2HPO
2 0.04
MgSO
4·7H
2O 0.04
MnSO
4 0.002
Water surplus (supplying 100%)
Cultivated 30 hours according to solid slant strains culture method in 32 ℃ incubator a temperature maintenance, obtaining bacterial classification is preserved under 2 ℃ condition, stand-by.
Step 2.-shake a bottle spawn culture
As the bacterial classification source, the prescription of the substratum of use is from step slant strains 1. in use:
Composition
Weight %
Sucrose 3
Soybean cake powder 3
Cottonseed cake powder 2
Peptone 0.2
Extractum carnis 0.5
Yeast powder 0.5
NaCl 0.2
(NH
4)
2SO
4 0.5
K
2HPO
2 0.04
MgSO
4·7H
2O 0.03
MnSO
4 0.002
Water surplus (supplying 100%)
Use the shaking table of a routine, the triangular flask of 50 500ml is housed on the shaking table, the 100ml culture medium solution is housed in each triangular flask, inoculate by inoculum size 3%.Shaking speed is 200rpm, cultivates on shaking table 30 hours, and the gemma number of the Bacillus licheniformis CGMCC No.1009 that records with the microscopy method is about 1,300,000,000/ml, then finish cultivation, as the bacterial classification source of next procedure.
3. a-500L seeding tank fermentation of step
Use, is inoculated by inoculum size 1% as the bacterial classification source from step strain liquid 2., and the prescription of the substratum of use is:
Composition
Weight %
Sucrose 5
Soybean cake powder 3
Cottonseed cake powder 2
Peptone 0.3
Extractum carnis 0.5
Yeast powder 0.5
NaCl 0.5
(NH
4)
2SO
4 0.5
K
2HPO
2 0.04
MgSO
4·7H
2O 0.03
MnSO
4 0.002
Water surplus (supplying 100%)
Fermentating controling condition is: tank pressure 0.2Kg/cm
2, air flow is 50m
3/ hour, in fermentor tank, to cultivate 48 hours, the gemma number that records Bacillus licheniformis with the microscopy method is about 4,000,000,000/ml, then finish to cultivate, as the bacterial classification source of next procedure.
Step is the b-20000L ferment tank 3.
Use from step 3. the strain liquid of a inoculate by inoculum size 1% as the bacterial classification source, 3. a is identical with step for the prescription of the substratum of use.Tank pressure 0.2Kg/cm
2, air flow is 300m
3/ hour, in back 12 hours of inoculation beginning, microscopy is carried out in sampling in per 4 hours, measures gemma quantity, and with the concentration of Waters224 type high performance liquid chromatograph (U.S.) quantitative assay zeatin, isopentenyl gland purine and ribosylzeatin.When fermentation culture to 48 hour, the gemma number that records is about 4,500,000,000/ml, and zeatin content is 40.6ppm (40.6 μ g/ml), and isopentenyl gland purine content is 2.7ppm, and ribosylzeatin content is 3.8ppm.So stop fermentation and put jar.
Step 4.-thin film concentration
With the thin film concentration method with step 3. institute's fermented liquid that obtains concentrate 5 times.
Step 5.-regulate to handle
In the concentrated broth that 4. step obtains, add dilute hydrochloric acid so that its pH value is adjusted to pH5, add 2% emulsifying agent 0203B then, stir.Thereby obtained the concentrated solution (the about 200ppm of zeatin) of plant-growth regulator of the present invention.
Application examples 1
The concentrated solution of the plant growth promoter that use obtains in embodiment 1, it is diluted 500 times, 1000 times, 1500 times respectively after, be sprayed on the leaf of cucumber plant by the amount of application of 50Kg diluent/mu, sprayed once every 7 days, add up its output.In addition, in contrast, use the plant growth promoter of purchasing on the market " cloud is big by 120 ", after 2000 times of its explanation dilutions, also the amount of application by 50Kg diluent/mu is sprayed on the leaf of cucumber plant, adds up its output.In addition, also with the cucumber of not spraying any medicament as blank.With obtain that the results are shown in Table 1.
Table 1 plant growth promoter is to the influence of cucumber yield
| The plant growth promoter kind | Extension rate | Cell production (kilogram) | Stimulation ratio (%) |
| Concentrated solution of the present invention | 500 | 85.25 | 34.8 |
| Concentrated solution of the present invention | 1000 | 79.25 | 25.3 |
| Concentrated solution of the present invention | 1500 | 75.75 | 19.8 |
| Commercially available cloud is big by 120 | 2000 | 75.50 | 19.4 |
| Blank | - | 63.25 | 0 |
As can be seen from Table 1, plant growth promoter of the present invention has clearly effect of increasing production to cucumber.Should illustrate, though commercially available cloud big 120 when diluting 1500 times has essentially identical effect of increasing production with plant growth promoter of the present invention in dilution 2000 times the time.But cloud big 120 is cloud tongue elements of a kind of artificial chemosynthesis, and plant-growth regulator of the present invention is the crude substance zeatin.
Application examples 2
The Xinjiang native teacher of the farming institute of agricultural sciences uses plant-growth regulator of the present invention that the cotton field is tested in the plant protection chamber.During test, use the concentrated solution of the plant-growth regulator that obtains among the embodiment 1, it is diluted 500 times, 1000 times and 2000 times respectively after, be sprayed on the leaf of cotton plants by the amount of application of 50Kg diluent/mu, sprayed once every 10 days.The result shows that conditioning agent of the present invention has than obvious facilitation the growth impetus of cotton.The plant height of twice investigation, the cotton plants of using above-mentioned three kinds of diluents all is higher than contrast, does not occur the overgrowing phenomenon again.Flower bud number, bell number and peach number all have raising in various degree, and can accelerate the growing process of cotton, and cotton is had very big production-increasing function.Institute obtains and the results are shown in the table 2.
Table 2 plant-growth regulator of the present invention is to the influence of output of cotton
| Extension rate | Peach number/rice 2 | Output Kg/ mu | Rate of increase (%) |
| 500 | 131.8 | 438 | 16.5 |
| 1000 | 134.4 | 448 | 19.1 |
| 2000 | 137.2 | 391 | 4.0 |
| Blank | 112.4 | 376 | 0 |
Application examples 3
Plant-growth regulator of the present invention is applied to food crop such as corn, wheat, paddy rice, jowar respectively according to Dilution ratio and the application process identical with application examples 1; Farm crop such as oil crops such as soybean, peanut and vegetables, melon and fruit, flowers, tobacco, fruit tree are all received the effect of increasing production of degree difference (20～30%).
Claims (9)
1. Bacillus licheniformis that is used to produce phytocytomine, it is characterized in that, it is Bacillus licheniformis CGMCC No.1009 (Bacillus Licheniformis), and said phytocytomine is zeatin, isopentenyl gland purine and ribosylzeatin.
2. a plant growth regulators is characterized in that, wherein contains Bacillus licheniformis CGMCC No.1009 and as zeatin, isopentenyl gland purine and the ribosylzeatin of its meta-bolites.
3. the production method of a plant growth regulators, comprise: 1. produce spawn culture, 2. shake a bottle spawn culture, 3. ferment tank cultivate, 4. thin film concentration, 5. five steps of emulsification, it is characterized in that, use Bacillus licheniformis CGMCC No.1009 as the bacterial classification source in 1. in step; Step 1., 2., the temperature that adopts in 3. the culturing process is 32 ℃, the pH value is 7 ± 0.1; The pH value of the concentrated solution that 4. step is obtained in 5. in step is adjusted to 5, and adds emulsifying agent to the particulate in the concentrated solution and can stably suspend.
4. the production method of plant-growth regulator as claimed in claim 3 is characterized in that, 3. wherein said step is divided into the 3. medium-sized ferment tank cultivation of a again; 3. two steps of b large fermentation tank fermentation culture.
5. the production method of plant-growth regulator as claimed in claim 3, it is characterized in that the weight percent of each composition of substratum that uses is: sucrose 1～3%, peptone 0.1～0.5%, yeast powder 0.3～0.5%, extractum carnis 0.3～0.7%, agar 1～3%, (NH when 1. production spawn culture of step
4)
2SO
40.3 K～0.5%,
2HPO
20.03 MgSO～0.05%,
47H
2O 0.02～0.05%, MnSO
40.001～0.003%, water supplies 100%;
At the step weight percent that shakes each composition of substratum that uses when bottle spawn culture and step ferment tank is 3. cultivated 2. be: sucrose 2～6%, soybean cake powder 2～5%, cottonseed cake powder 1～3%, peptone 0.1～0.5%, extractum carnis 0.3～0.7%, yeast powder 0.3～0.7%, NaCl0.1～0.6%, (NH
4)
2SO
40.3 K～0.7%,
2HPO
20.02 MgSO～0.05%,
47H
2O0.02～0.05%, MnSO
40.001～0.003%, water supplies 100%.
6. the production method of plant growth promoter as claimed in claim 3 is characterized in that, the emulsifying agent that adds in 5. in step is 0203B, and its consumption is 1～2 weight % of plant-growth regulator.
7. the production method of plant-growth regulator as claimed in claim 3 is characterized in that, 1. step carries out in incubator, incubation time 24～36 hours, and the slant strains that obtains is preserved under 0～4 ℃ temperature, uses in 1 year; 2. step carries out on shaking table, and shaking speed is 150～300rpm, finishes when the gemma number that records with microscopy reaches 10～2,000,000,000/ml to cultivate; 3. step carries out in the industrial fermentation jar, the air air flow be 15～100 tank volume/hour, the gemma number that records with microscopy reach 4,000,000,000/more than the ml, the content of zeatin reaches 40 μ g/ml and finishes cultivation when above simultaneously, put jar, and then carry out step 4. with step 5..
8. the described plant-growth regulator of claim 2 is used to promote the purposes of crop growth and volume increase.
9. plant-growth regulator as claimed in claim 8 is used to promote the purposes of crop growth and volume increase, it is characterized in that, wherein said farm crop are food crop, oil crops, vegetables, melon and fruit, cotton, flowers, tobacco and fruit tree.
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|---|---|---|---|
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|---|---|---|---|
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| CN101891548B (en) * | 2010-07-21 | 2014-06-18 | 神州汉邦（北京)生物技术有限公司 | Multiple-effect microbial fertilizer prepared by three strains of bacillus licheniformis |
| CN102732523B (en) * | 2012-07-02 | 2014-05-14 | 中国科学院化学研究所 | Aptamer for identifying zeatin through specifity, and screening method and application of aptamer |
| CN102765809B (en) * | 2012-07-05 | 2013-10-16 | 昆明豪原特自控有限公司 | Biological wetland forming growth promoter and preparation method thereof |
| CN103329939A (en) * | 2013-07-10 | 2013-10-02 | 唐睿 | Compound fermentation formula and technology of compound amino acid purine mother liquor Niannianle |
| CN104041521B (en) * | 2014-03-26 | 2017-01-04 | 贵州省烟草科学研究院 | The application in field produces of water-retaining agent and Nicotiana tabacum L. Promoting bacteria |
| UY36334A (en) * | 2014-12-29 | 2017-04-28 | Fmc Corp | COMPOSITIONS OF BACILLUS LICHENIFORMIS RT1184 AND USES METHODS TO BENEFIT THE GROWTH OF PLANTS |
| AR102123A1 (en) * | 2014-12-29 | 2017-02-08 | Fmc Corp | SEED MATRICES THAT INCLUDE MICROORGANISMS OF BACILLUS SPP. TO BENEFIT THE GROWTH OF THE PLANT |
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