CN1446464A - Method for preparing high efficiency biological weed control bacterial agent and its usage - Google Patents
Method for preparing high efficiency biological weed control bacterial agent and its usage Download PDFInfo
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Abstract
A process for preparing the efficient bacterial herbicide includes sampling the soil near the root of weeds such as crabgrass herb, caper euphorbia, cassia, etc. or their stem or leaves, separating with NPC culture medium, and the chlorella pyrenoidosa and glutamine synthetase depressants, and screening the bacterial strains with high herbiciding activity and broad spectrum. Its advantage is high herbiciding effect.
Description
Technical field
The present invention relates to the Preparation method and use of pathogenic mutation (the Xanthomonas campestris pv.retroflexus) microbial inoculum of a kind of biological weed control bacterial strain xanthomonas campestris Amaranthus retroflexus.
Background technology
Chemical herbicide can effectively be controlled many malignant weeds, but its a large amount of uses have also caused a series of environmental problems, and the development difficulty of new chemical herbicide is big, success rate is low, expensive height.By contrast, microbial herbicide low toxicity, easily degraded, Environmental compatibility are good, have huge development potentiality.
The research of microbial herbicide starts from the sixties in last century, successively releases a collection of commercial mycoherbicide, and academy of agricultural sciences, China Shandong has developed the culture control soybean field dodder of " Shandong is protected No. one " dodder Colletotrichum gloeosporiodes; These mycoherbicides all are spore preparation, and action condition is strict, require too highly in that batch process, prescription, storage etc. are technical, are not widely accepted, and do not produce remarkable social benefit and economic benefit.The early 1990s, people turned to bacterium with goal in research, filtered out inhibited bacterium from the microorganism species of weeds root soil, by cultivating born of the same parents' exotoxin that acquisition is caused a disease the host.Because bacterium is short than the conk cycle, zymotechnique is simple, production procedure is easy to control, its born of the same parents' exotoxin to environmental condition require not resemble the fungal spore strictness, easily by soil degrading, have the good application development prospect, become a big focus of microbial herbicide development.
Root bacterium with herbicide effect mainly is pseudomonas (Pseudomonas), Erwinia (Erwinia), the Xanthomonas (Xanthomonas) of Gram-negative bacteria, and what wherein study at most is pseudomonas.The phytotoxin phaseolotoxin that Pseudomonas syringae pv.phaseolicola is produced can make the blade of elegant jessamine (Kudzu) chlorotic illness occur, produces local necrosis; In a single day this toxin enters plant root and will infect to the branch end, cause dwarfing, the chlorosis of plant, serious causes the plant leaf blade necrosis.Gurusiddaiah.S will cause thick toxin behind the Pseudomonas fluorescensstrain D7 bacterial strain fermentation liquor preliminary purification of disease in order to control cheatgrass brome or the like.Johnson B.J. studies Xanthomonas campestris at first as biological weed killer, and he filters out many strains has the edible wild herbs Xanthomonas campestris (Xanthomonas Campestris pv.poannua) of activity of weeding to be used for controlling the annual annual bluegrass (Poa annua L.) on lawn, Bermuda.Mainly be applicable to non-irrigated greenery patchess such as lawn, peanut ground, potato field, road both sides, golf course.
Summary of the invention
The objective of the invention is to based on plant ecology principle, separate the vegetable active bacterium that obtains bacteriocinogeny from rhizosphere soil or plant tissue; And provide this preparation method who the field main weeds is had the biologically active microbial inoculum of the effect of preventing and kill off.Therefore, the starting point of the present invention is to screen has the effect of preventing and kill off and to the bacterial strain of most of turfgrass and nursery stock safety, it can prevent and treat multiple weeds in field to the field main weeds, and its activity of weeding is strong, broad weed-killing spectrum, selectivity are good.Through a large amount of screening operations, from rhizosphere bacteria, filter out the mutation of causing a disease of the bacterial strain xanthomonas campestris Amaranthus retroflexus with higher activity of weeding, clear and definite its target weeds, kill careless spectrum, using dosage and range of application.
The preparation method of biological weed control microbial inoculum of the present invention, concrete steps are as follows:
1) from weeds rhizosphere 5-15cm soil such as Amaranthus retroflexus, lady's-grass, lamb's-quarters, moleplant seed, Cassia tora, terrestrial stem, blade sampling, rinsing is 1-3 time in the 200ml sterile water, 400-600rpm shakes a bottle 10min in the input 100ml 0.01-2.0% Tween-80 solution, with sterile phosphate buffer solution (5-20Mm K
3PO
4-KH
2PO
40.05-2MNaCl, PH6.4-8.2) make 10-20 times of serial dilution, get 0.1-0.3ml and coat the NPC culture medium flat plate, cultivate the 20-30h observed result for 20-32 ℃, select coloured bacterium colony, be stored on the nutrient agar, the single bacterium colony that just sifts out is further separated purifying: 1. bacteriostatic test, connect ring bacterium to be measured with transfer needle, percutaneous puncture-inoculation is to the Escherichia coli flat board, and 20-32 ℃ of incubator cultivated 20-30h, observes antibacterial spot; 2. chlorella pyrenoidosa (Chlorella pyrenoidosa) growth inhibition experiment is inoculated into algae liquid in the conical flask, and initial concentration is 8 * 10
3-8 * 10
6Individual cell/ml; Handle to add liquid to be measured, contrast does not add, and 20-30 ℃, 3000-6000lux illuminance continuous light and 100-200rpm rotational oscillation are cultivated 2-7d, are reference with the culture fluid, measure light absorption value (light path 1cm) under maximum absorption wavelength 680nm, survey inhibiting rate; 3. glutamine synthetase inhibitor model discrimination, be coated with bacillus subtilis bacterial suspension flat board not adding glutamine and added on the conventional medium of glutamine that concentration is 0.05-10% respectively, observe fungistatic effect, it is fast therefrom to select colony growth, good antimicrobial effect, the bacterial strain that biologically active is high.
2) bacterial strain carries out slant culture in conventional solid culture medium, and rejuvenation is cultivated 9-24h for 25-30 ℃; Carry out seed culture then, contain the liquid nutrient medium of 2/3 volume in the 400ml triangular flask, the prescription of liquid nutrient medium is: beef extract 0.5-2.2%, peptone 0.2-4%, glucose 0.2-4%, NH4H2PO40.05-0.5%, NaCl 0.1-2.0% supplies volume with water; Accent initial p H6.0-7.4, at the 121-125 ℃ of 20-30min that sterilizes down, the cooling back is inoculated with 1% inoculum concentration, cultivates 12-32h for 25-30 ℃.
3) liquid nutrient medium of Sheng 2/3 volume in the fermentation tank, the prescription of liquid nutrient medium are with the prescription of seed liquid nutrient medium, and with the inoculum concentration inoculation of 6-12%, throughput is 1: 0.6-1.5 (V: V), cultivate 54-120h for 25-30 ℃; Bacteria containing amount is 10 in the fermented liquid
5-9Cfu/ml; Add auxiliary agent, auxiliary agent is 0.5-5% Tween 80 or ammonia silicon, obtains the weeding microbial inoculum; Directly be packaged into product.
The above-mentioned first step is selected, and to have the mutation of causing a disease of high-performance bio activity of weeding bacterial strain xanthomonas campestris Amaranthus retroflexus be 15-30mm to colibacillary inhibition zone, it is strong that glutamine synthelase is suppressed ability, and the zymotic fluid dilution 10-30 that is suppressed at of chlorella pyrenoidosa is doubly had 60-90% down.
The population characteristic of the pathogenic mutation of xanthomonas campestris Amaranthus retroflexus: bacterium colony is rounded on solid plate, full edge, and smooth surface is moistening, and is translucent, yellow.Colony diameter is 5-6mm/24h; Do not have mycoderm during static cultivation in the liquid medium within and form, cytotostatic, bacterium liquid become muddy also has precipitation to produce.
Individual morphology: cell is a rod-short, 1.5-2.0 * 0.5-1.0um, and no gemma, polar flagella, motion, leather Albert'stain Albert feminine gender has the class fat granule.
Physicochemical characteristics: the catalase reaction aerogenesis, aerobic, fixing nitrogen can not utilize one-carbon compound as unique carbon source.Growth temperature is 8-50 ℃, and optimum growth temperature is 25-35 ℃; PH5-9 can both grow, optimal pH 6-7.5.Can produce non-water-soluble carotenoid, the oxydase reaction feminine gender can not be reduced nitrate.Energy hydrolyzation of glucose, gelatin can not hydrolyzed starches.
According to above-mentioned feature, the Xanthomonas (Xanthomonas) of this Pseudomonas in the 8th edition " uncle Jie Shi determinative bacteriology handbook ", this bacterium is in China Committee for Culture Collection of Microorganisms of the depositary institution common micro-organisms center preservation of Patent Office of the People's Republic of China's appointment, and deposit number is: CGMCC NO.0902.The cultural characteristic of this bacterial strain, personal feature and physiological and biochemical property see Table 1,2.
The application of high-performance bio weeding microbial inoculum of the present invention
High-performance bio weeding microbial inoculum of the present invention is used for turfgrass and woody nursery stock and draft nursery stock and prevents and kill off broad leaved weed and grassy weed.
Said turfgrass is Festuca Arundinacea, Bermuda grass, dewdrop grass, perennial ryegrass, Korea lawn grass etc.Nursery stock be fragrant camphor tree, cdear, southern magnolia, sweet osmanthus, cryptomeria, Chinese scholar tree, Chinese littleleaf box, glossy privet, Fructus Manglietiae insignis, gold leaf with a smile, Chinese wistaria, reach the clouds etc.Broad leaved weed and grassy weed are mainly Amaranthus retroflexus, kitchen garden, thorn amaranth, shepherd's purse, barnyard grass grass, green foxtail, eleusine indica, lady's-grass, annual bluegrass, amur foxtail, two fringe province barnyard grass, lamb's-quarters, Siberian cocklebur, shepherd's purse, clearvers, Veronica, field bindweed, alternanthera philoxeroides, chrysanthemum punt-pole, cotton wool sorrel Liao, Hairy Bittercress, the rhizome of nutgrass flatsedge etc.
Using method, dosage and period: adopt the method dispenser of spraying, using dosage is 500-1000L/ha; Operating period is best in weed germination to the five leaf phase, the general first half of the year 3, April, the second half year 9, October; Microbial inoculum is done once also can use as secondary, for the second time dispenser one to four week after the dispenser first time.
Biological weed control microbial inoculum of the present invention shows that through temperature, humidity and illumination condition test this microbial inoculum is comparatively loose to environment requirement, and temperature is at 10-35, and humidity is more than 20%, pathogenic all good to the target weeds; Generally after dispenser, showed in 1 week pathogenicly, can continue until more than 4 weeks.Do not have residually, do not have pathogenicly, environmentally safe, microbial inoculum also can play the effect of enrichment edaphon.This microbial inoculum also is applicable to large-scale production simultaneously, and effective strain is simple, and technology is simple, and is with low cost, easy to use.
Description of drawings
Fig. 1 is the process chart that the pathogenic mutation of xanthomonas campestris Amaranthus retroflexus that separation obtains is prepared microbial inoculum.
Embodiment
Below further specify the preparation method of biological weed control microbial inoculum by example, concrete steps are as follows:
1) uses from Amaranthus retroflexus weeds rhizosphere 10cm soil, terrestrial stem, blade sampling, remove the unnecessary soil of weeds root, rinsing is 2 times in the 200ml sterile water, and 500rpm shakes a bottle 10min in the input 100ml 1.0% Tween-80 solution, with sterile phosphate buffer solution (6MmK
3PO
4-KH
2PO
4, 0.2MNaCl PH7.2) makes 10 times of serial dilutions, get 0.1ml and coat the NPC culture medium flat plate, the NPC culture medium flat plate refers to add penicillin, ovobiocin, cycloheximide in the DavisShi minimal medium, cultivates the 22h observed result for 24 ℃, select coloured bacterium colony, be stored on the nutrient agar.The single bacterium colony that just sifts out is further separated purifying: 1. bacteriostatic test, connect ring bacterium to be measured with transfer needle, percutaneous puncture-inoculation is to the Escherichia coli flat board, and 24 ℃ of incubators are cultivated 22h, observe antibacterial spot, and used medium is conventional medium.2. chlorella pyrenoidosa (Chlorellapyrenoidosa) growth inhibition experiment is inoculated into algae liquid in the conical flask, and initial concentration is 8 * 10
3Individual cell/ml.Handle to add fermented liquid to be measured, contrast does not add, and 22 ℃, 3000lux illuminance continuous light and 100rpm rotational oscillation are cultivated 3d, are reference with the culture fluid, measure light absorption value (light path 1cm) under maximum absorption wavelength 680nm, survey inhibiting rate.3. glutamine synthetase inhibitor model discrimination is coated with bacillus subtilis bacterial suspension flat board not adding glutamine and added on the conventional medium that concentration is 2% glutamine respectively, observes fungistatic effect.It is fast therefrom to select colony growth, good antimicrobial effect, the bacterial strain that biologically active is high.The mutation of causing a disease of selected bacterial strain xanthomonas campestris Amaranthus retroflexus is 23mm to colibacillary inhibition zone, and it is strong that glutamine synthelase is suppressed ability, and being suppressed under 30 times of the zymotic fluid dilutions of chlorella pyrenoidosa had more than 70%.
2) bacterial strain carries out slant culture in conventional solid culture medium, and rejuvenation is cultivated 22h for 26 ℃; Carry out seed culture then, contain the liquid nutrient medium of 2/3 volume in the 400ml triangular flask, the prescription of liquid nutrient medium is: beef extract 0.5%, and peptone 3%, glucose 2%, NH4H2PO4 0.05%, and NaCl 1.5%, supplies volume with water; Accent initial p H7.4, at the 121-125 ℃ of 20min that sterilizes down, the cooling back is inoculated with 1% inoculum concentration, cultivates 28h for 26 ℃.
3) liquid nutrient medium of Sheng 2/3 volume in the fermentation tank, the prescription of liquid nutrient medium is with the prescription of seed liquid nutrient medium, and the inoculum concentration with 6% is inoculated, and throughput is 1: 0.7 (V: V), cultivate 96h for 26 ℃; Bacteria containing amount is 10 in the fermented liquid
7Cfu/ml; Add auxiliary agent, auxiliary agent is 4% Tween 80 or ammonia silicon, obtains the weeding microbial inoculum; Directly be packaged into product.
The application example of high-performance bio weeding microbial inoculum is as follows:
Embodiment 1: Amaranthus retroflexus
Showing money or valuables one carries unintentionally to sprout to 4 leaf periods at the Amaranthus retroflexus seed sprays microbial inoculum, and dosage shows preventive effect (seeing Table 3) preferably at 500L/ha.New leaf withering behind the 1d, Lao Ye begins to wilt behind the 2d, and this withers 4d posterior lobe subbase, and preventive effect reaches more than 90%.Preventive effect slightly descends after 3 weeks, and the part cauline leaf of Amaranthus retroflexus begins to recover, but growing way is poor, protected plant is not constituted influence.
Embodiment 2: Cassia tora
Show money or valuables one carries unintentionally to sprout at the Cassia tora seed and take turns leaf period to the 4th and spray microbial inoculum, dosage shows preventive effect preferably at 500L/ha.Young leaves begins to wilt behind the 2d, and 3d rear section Lao Ye begins to wilt, and this withers 7d posterior lobe subbase, and preventive effect reaches more than 85%.Preventive effect descends to some extent after 3 weeks, and the small part cauline leaf of Cassia tora begins to recover, but the growing way extreme difference does not constitute influence to protected plant.
Embodiment 3: little lamb's-quarters
Showing money or valuables one carries unintentionally to sprout to 4 leaf periods at little lamb's-quarters seed sprays microbial inoculum, and dosage is at 500L/ha, and preventive effect is good.This withers 5d posterior lobe subbase, and preventive effect reaches more than 85%, and preventive effect slightly descends after 3 weeks, plants following but be controlled in economic threshold.
Embodiment 4: annual bluegrass
Showing money or valuables one carries unintentionally to sprout to 3 leaf phases at the annual bluegrass seed sprays microbial inoculum, and dosage is at 500L/ha, and preventive effect is better.Most of leaf is withered behind the 4d, and preventive effect reaches 75%.Preventive effect descends to some extent after 2 weeks.
Embodiment 5: lady's-grass
Showing money or valuables one carries unintentionally to sprout to 4 leaf periods at the lady's-grass seed sprays microbial inoculum, and dosage shows preventive effect preferably at 500L/ha.New leaf withering behind the 1d, old leaf withering behind the 2d, this withers 4d posterior lobe subbase, and preventive effect reaches more than 85%, and preventive effect begins to descend after 3 weeks.
Embodiment 6: Festuca Arundinacea
Spray microbial inoculum at Festuca Arundinacea 2 leaves that sprout after phase, dosage shows safety preferably (seeing Table 4) at 900L/ha.The above no poisoning of 3 leaf phases, 2 the leaf phase plant slightly downgrade, both can recover after 1 week, safer under the low dosage.So it is safer that 2 leaf after date dispensers are grown at Festuca Arundinacea in the field.
Embodiment 7: gold leaf with a smile
Grow to after gold leaf sprouts with a smile and spray microbial inoculum more than the 4cm, dosage shows safety preferably (seeing Table 5) at 900L/ha.Spraying growth at 3-4cm has suffered inhibition, but very fast recovery.So field dispenser safety after gold leaf grows to the 4.5cm height with a smile.
Embodiment 8: Fructus Manglietiae insignis
Grow to after Fructus Manglietiae insignis sprouts and spray microbial inoculum more than the 5cm, dosage is at 900L/ha, and safety is better.Pathogenic mutation cultural characteristic of table 1. xanthomonas campestris Amaranthus retroflexus and personal feature
| The solid plate colony characteristics | Size | ????5-6mm |
| Shape | Circular | |
| Surface texture | Smooth moistening | |
| The edge | Full edge | |
| Optical signature | Translucent | |
| The lawn color | Yellow | |
| Chromogenesis | Have | |
| Liquid is supported the body training | Mycoderm | Do not have |
| Muddy | Have | |
| Precipitation | Have | |
| Personal feature | Wide (um) | ????0.5-1.0 |
| Long (um) | ????1.5-2.0 | |
| The leather Albert'stain Albert | Negative | |
| Motility | Have | |
| Flagellum | Extremely give birth to＞1 | |
| The dyeing of class fat granule | Have |
The table 2. xanthomonas campestris Amaranthus retroflexus mutation physiological and biochemical property that causes a disease
| Catalase | ????+ |
| Oxidase | ????- |
| The glucose oxidase fermentation test | ????+ |
| Gelatin hydrolysis | ????+ |
| The starch hydrolysis | ????- |
| The L-valine | ????+ |
| Beta-alanine | ????+ |
| The DL-arginine | ????+ |
| Nitrate reduction | ????- |
| 41 ℃ of growths | ????+ |
| The mensuration of arginine dihydrolase | ????- |
| The generation of fluorchrome | ????- |
Table 3 microbial inoculum is to different leaf age phase weeds preventive effect (fresh weight inhibiting rate %)
| The weeds title | Leaf age | ||||
| ????1 | ????2 | ????3 | ????4 | ????5 | |
| Amaranthus retroflexus | ????98.1 | ????90.5 | ????86.6 | ????82.4 | ????45.2 |
| Cassia tora | ????100.0 | ????93.4 | ????84.1 | ????81.3 | ????50.7 |
| Little lamb's-quarters | ????100.0 | ????94.3 | ????88.6 | ????84.2 | ????67.8 |
| Knotweed | ????95.6 | ????86.4 | ????80.1 | ????71.8 | ????39.6 |
| Before the car | ????98.2 | ????91.4 | ????86.3 | ????69.4 | ????41.8 |
| Garden sorrel | ????94.0 | ????88.7 | ????79.4 | ????71.4 | ????52.6 |
| Annual bluegrass | ????95.0 | ????87.7 | ????78.5 | ????63.4 | ????41.7 |
| Lady's-grass | ????100.0 | ????96.3 | ????90.5 | ????86.2 | ????66.4 |
| Moleplant seed | ????94.7 | ????86.7 | ????74.5 | ????65.7 | ????44.9 |
| Eleusine indica | ????90.4 | ????80.0 | ????67.5 | ????53.3 | ????37.7 |
Table 4 microbial inoculum is to the poisoning situation (fresh weight inhibiting rate %) of different leaf age phase turfgrass
| The turfgrass title | Leaf age | |||
| ????1 | ????2 | ????3 | ????4 | |
| Festuca Arundinacea | ??45.7 | ??15.6 | ????6.3 | ????3.4 |
| Bermuda grass | ??51.3 | ??20.4 | ????5.5 | ????0 |
| Dewdrop grass | ??66.7 | ??17.3 | ????9.1 | ????3.6 |
| Perennial ryegrass | ??53.4 | ??19.3 | ???-1.3 | ????2.7 |
Table 5 microbial inoculum is to the poisoning situation (fresh weight inhibiting rate %) of different plant height nursery stocks
| The turfgrass title | Plant height (cm) | |||
| ????2 | ????3 | ????4 | ????5 | |
| Gold leaf with a smile | ??55.7 | ??31.6 | ???7.8 | ???4.4 |
| Mount emei with a smile | ??56.3 | ??40.4 | ???22.5 | ???4.9 |
| Fructus Manglietiae insignis | ??62.7 | ??47.6 | ???27.6 | ???6.6 |
Claims (5)
1. the preparation method of a high-performance bio weeding microbial inoculum is characterized in that may further comprise the steps:
1) from weeds rhizosphere 5-15cm soil such as Amaranthus retroflexus, lady's-grass, lamb's-quarters, moleplant seed, Cassia tora, terrestrial stem, blade sampling, rinsing is 1-3 time in the 200ml sterile water, 400-600rpm shakes a bottle 10min in the input 100ml 0.01-2.0% Tween-80 solution, with sterile phosphate buffer solution (5-20Mm K
3PO
4-KH
2PO
40.05-2MNaCl, PH6.4-8.2) make 10-20 times of serial dilution, get 0.1-0.3ml and coat the NPC culture medium flat plate, cultivate the 20-30h observed result for 20-32 ℃, select coloured bacterium colony, be stored on the nutrient agar, the single bacterium colony that just sifts out is further separated purifying: 1. bacteriostatic test, connect ring bacterium to be measured with transfer needle, percutaneous puncture-inoculation is to the Escherichia coli flat board, and 20-32 ℃ of incubator cultivated 20-30h, observes antibacterial spot; 2. chlorella pyrenoidosa (Chlorella pyrenoidosa) growth inhibition experiment is inoculated into algae liquid in the conical flask, and initial concentration is 8 * 10
3-8 * 10
6Individual cell/ml; Handle to add liquid to be measured, contrast does not add, and 20-30 ℃, 3000-6000lux illuminance continuous light and 100-200rpm rotational oscillation are cultivated 2-7d, are reference with the culture fluid, measure light absorption value (light path 1cm) under maximum absorption wavelength 680nm, survey inhibiting rate; 3. glutamine synthetase inhibitor model discrimination, be coated with bacillus subtilis bacterial suspension flat board not adding glutamine and added on the conventional medium of glutamine that concentration is 0.05-10% respectively, observe fungistatic effect, it is fast therefrom to select colony growth, good antimicrobial effect, the bacterial strain that biologically active is high.
2) bacterial strain carries out slant culture in conventional solid culture medium, and rejuvenation is cultivated 9-24h for 25-30 ℃; Carry out seed culture then, contain the liquid nutrient medium of 2/3 volume in the 400ml triangular flask, the prescription of liquid nutrient medium is: beef extract 0.5-2.2%, peptone 0.2-4%, glucose 0.2-4%, NH
4H
2PO
40.05-0.5%, NaCl 0.1-2.0% supplies volume with water; Accent initial p H6.0-7.4, at the 121-125 ℃ of 20-30min that sterilizes down, the cooling back is inoculated with 1% inoculum concentration, cultivates 12-32h for 25-30 ℃.
3) liquid nutrient medium of Sheng 2/3 volume in the fermentation tank, the prescription of liquid nutrient medium are with the prescription of seed liquid nutrient medium, and with the inoculum concentration inoculation of 6-12%, throughput is 1: 0.6-1.5 (V: V), cultivate 54-120h for 25-30 ℃; Bacteria containing amount is 10 in the fermented liquid
5-9Cfu/ml; Add auxiliary agent, auxiliary agent is 0.5-5% Tween 80 or ammonia silicon, obtains the weeding microbial inoculum;
2. the application of high-performance bio weeding microbial inoculum according to claim 1 is characterized in that this microbial inoculum is used for turfgrass and woody nursery stock and draft nursery stock and prevents and kill off broad leaved weed and grassy weed.
3. the application of high-performance bio weeding microbial inoculum according to claim 2 is characterized in that said turfgrass is Festuca Arundinacea, Bermuda grass, dewdrop grass, perennial ryegrass, Korea lawn grass etc.
4. the application of high-performance bio weeding microbial inoculum according to claim 2, it is characterized in that said nursery stock be fragrant camphor tree, cdear, southern magnolia, sweet osmanthus, cryptomeria, Chinese scholar tree, Chinese littleleaf box, glossy privet, Fructus Manglietiae insignis, gold leaf with a smile, Chinese wistaria, reach the clouds etc.
5. the application of high-performance bio weeding microbial inoculum according to claim 2 is characterized in that said broad leaved weed and grassy weed are mainly Amaranthus retroflexus, kitchen garden, thorn amaranth, shepherd's purse, barnyard grass grass, green foxtail, eleusine indica, lady's-grass, annual bluegrass, amur foxtail, two fringe province barnyard grass, lamb's-quarters, Siberian cocklebur, shepherd's purse, clearvers, Veronica, field bindweed, alternanthera philoxeroides, chrysanthemum punt-pole, cotton wool sorrel Liao, Hairy Bittercress, the rhizome of nutgrass flatsedge etc.
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| CN100565187C (en) * | 2007-09-06 | 2009-12-02 | 浙江工业大学 | A kind of high-throughout 96 orifice plate screening techniques of herbicide that are used for |
| CN101611707B (en) * | 2009-08-12 | 2014-07-16 | 海南利蒙特生物农药有限公司 | Bacillus subtilis oil suspending agent and preparation method thereof |
| CN102206604A (en) * | 2011-04-28 | 2011-10-05 | 南京大学 | Enterobacter agglomerans strain and application thereof in weeding |
| CN109090144A (en) * | 2018-10-17 | 2018-12-28 | 马书文 | A kind of novel environmentally-friendly biological herbicide |
| CN110129062A (en) * | 2019-06-14 | 2019-08-16 | 阿尔格生命科学(江苏)有限公司 | A kind of algae competent cell soil conditioning agent producing process |
| CN110590593A (en) * | 2019-09-23 | 2019-12-20 | 青岛农业大学 | Phenolic acid amide derivative and preparation method and application thereof |
| CN110590593B (en) * | 2019-09-23 | 2022-10-18 | 青岛农业大学 | Phenolic acid amide derivative and preparation method and application thereof |
| CN114441491A (en) * | 2022-01-25 | 2022-05-06 | 河北科技大学 | Method for detecting atrazine biotoxicity by chlorella pyrenoidosa fluorescence |
| CN114657068A (en) * | 2022-04-11 | 2022-06-24 | 广西地源之本肥业有限公司 | Preparation method of Kangtu weed-inhibiting bacteria |
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