CN101632367B - Application of biopesticide tetramycin in preventing and controlling Magnaporthe grisea - Google Patents
Application of biopesticide tetramycin in preventing and controlling Magnaporthe grisea Download PDFInfo
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Abstract
The invention discloses novel application of biopesticide tetramycin generated by Streptomyces ahygroscopicus subsp.Wuzhouensis n.subsp in preventing and controlling Magnaporthe grisea. The tetramycin is compound agricultural antibiotic, comprises tetramycin A1 and A2, albonoursin and flagecidin, and has a rate of preventing and controlling the apple canker of over 95 percent. Indoor and field experiments prove that tetramycin diluent (the ratio of the tetramycin to water is 1: 999) has better effect of preventing and controlling the Magnaporthe grisea than 4 percent kasugamicin waterable powder, 75 percent tricyclazole waterable powder and 25 percent prochloraz missible oil, and has administration lower than positive control remarkably. The tetramycin used can reduce agricultural production cost, improves prevention effect, can be alternately used with other biopesticides, reduces the generation of resistance, has good effect of preventing and controlling the Magnaporthe grisea, and no problems such as environmental pollution, pesticide residue and the like, belongs to the environment-friendly biopesticide, and has good development and application prospects.
Description
Technical field
The present invention relates to a kind of by the not biopesticide of streptomyces hygroscopicus Wuzhou subspecies (Streptomyces ahygroscopicus subsp.Wuzhouensis n.subsp) generation---the application of tetramycin in the control rice blast.
Background technology
China is large agricultural country, plants cereal crops more than 100,000,000 hectares throughout the year, and cotton is more than 4,800,000 hectares, and oil crop are more than 1,400 ten thousand hectares, and vegetable melon and fruit needs the control area above 300,000,000 hectares more than 1,800 ten thousand hectares throughout the year.For prevention and elimination of disease and pests, guarantee the ordinary production of agricultural, for a long time, people use the chemical pesticide control damage by disease and insect in a large number.The use of chemical pesticide is played useful effect to guaranteeing the crops ordinary production; But along with using chemical pesticides for a long time and in a large number; Ecotope is done great damage; Like enzymic activity reduction in soil structure destruction, the soil, soil property and water body severe contamination or the like; Destroyed the whole ecological environment, poisonous persticide residue severe overweight in nearly all food such as grain, vegetables, fruit, the gourd, fruit and vegetable of edible residual chemical pesticide severe overweight causes person poultry poisoning's incident also to happen occasionally.
To China's WTO accession, developed country adjusts the technical threshold that has improved the product import one after another, and the influence that agricultural product receive stands in the breach.Only the agricultural byproducts loss that closed down by external " green barrier " is annual just up to 7,000,000,000 dollars in China.
Developing non-harmful biopesticide is the main path that solves existing agricultural chemicals and export of farm produce problem.Biopesticide belongs to environmentally friendly product; Use biopesticide; Not only can reduce the use of chemical pesticide and chemical fertilizer, realize the high-quality and safety production of agricultural product effectively, promote the economic value added of agricultural product; And can be agricultural products in China outlet and create favorable conditions, greatly strengthen the international competitiveness of agricultural products in China.China is advanced green agriculture industry development and developing rural economy, increases farmers' income, promotes the rural area prosperity to have important progradation.
Paddy rice is the main cereal crops of China, and cultivated area surpasses 2,800 ten thousand hectares at present, and annual production reaches more than 1.7 hundred million tons.Annual because the generation of damage by disease and insect all can cause the large-area underproduction, become the key factor that hinders rice high yield.Rice blast is the main disease of paddy rice, spreads soon, and it is serious to cause harm, and prevents and treats the bad meeting underproduction more than 50%.Now the main medicament of control rice blast has: tricyclazole wettable powder, execute rice spirit, Isoprothiolane missible oil, the disease of going out prestige colloidal suspending agent etc., but deposit all that medicament is residual, environmental pollution, be prone to produce problems such as resistance.
Tetramycin is as known compound; Be by streptomyces hygroscopicus Wuzhou subspecies (Streptomyces ahygroscopicussubsp.Wuzhouensis n.subsp) (" microorganism journal " 1988 not; 28 volume (1) 85-87 pages or leaves) the complex biological antibiotic that produces; Contain tetramycin A1, A2 and albonoursin and flagecidin; Unit prevents and treats the application studies such as apple canker, alternaria leaf spot of apple and graw mold of tomato tetramycin except that invention, does not see other unit also has tetramycin other new purposes as biopesticide relevant report as yet.
Summary of the invention
The present invention seeks to the defective to the prior art existence, the new purposes of tetramycin is provided, promptly tetramycin is as the application of biopesticide in the control rice blast.
The present invention relates to by streptomyces hygroscopicus Wuzhou subspecies (Streptomyces ahygroscopicus subsp.Wuzhouensis n.subsp) (" microorganism journal " 1988 not; (1) 8587 page of 28 volume), the tetramycin of generation is as the application of biopesticide in the control rice blast.
Tetramycin is a kind of complex biological antibiotic, contains tetramycin A1, A2 and albonoursin and flagecidin.According to molecular weight and some other physicochemical data that nuclear magnetic resonnance, infrared absorption spectroscopy, ultra-violet absorption spectrum and the mass spectrum of tetramycin A1 are confirmed, the molecular formula of deriving tetramycin A1 should be C
35H
52NO
13, its chemical structural formula is:
According to similar
tetramycin A1 isolation and identification method, inhale through nuclear magnetic resonnance, infrared absorption spectroscopy, ultraviolet
Receive molecular weight and some other physicochemical data that spectrum and mass spectrum are confirmed, derive tetramycin B chemical structural formula and be:
According to similar tetramycin A1 isolation and identification method,, derive tetramycin C chemical structural formula and be through molecular weight and the some other physicochemical data that nuclear magnetic resonnance, infrared absorption spectroscopy, ultra-violet absorption spectrum and mass spectrum are confirmed:
Following table is the tetramycin antimicrobial spectrum
| Indicator bacteria | The bacterial classification source | Connect resistance | Indicator bacteria | The bacterial classification source | Connect resistance |
| Bacillus sphaericus | In little institute | --- | Escherichia coli | Department of traditional Chinese medicine institute | --- |
| Pseudomonas fluorescens | In little institute | --- | Escherichia coli | In little institute | + |
| Gold-coloured staphylococci | In little institute | --- | Gold-coloured staphylococci | Department of traditional Chinese medicine institute | + |
| Bacillus cercus | In little institute | --- | Hay bacillus | In little institute | + |
| Bacteria erwinia | In little institute | --- | Brevibacterium flavum | In little institute | + |
| Pseudomonas aeruginosa | Department of traditional Chinese medicine institute | --- | Much's bacillus | In little institute | + |
| Sarcina lutea | Department of traditional Chinese medicine institute | ++ | The pan mould | In little institute | + |
| Saccharomyces cerevisiae | In little institute | ?++ | Fusarium graminearum | In little institute | + |
| Unusual Hansenula yeast | In little institute | ?--- | The beautiful broomcorn millet Helminthosporium sp. that belongs to | In little institute | - |
| Fission yeast | In little institute | ++ | Chain lattice spore is mould | In little institute | - |
| Rhodothece rubra | In little institute | ++ | The cotton anthracnose bacterium | In little institute | - |
| Ka Er Qi Shi uncle yeast | In little institute | ++ | The fusarium graminaria bacterium | In little institute | + |
| Rhodotorula glutinis | In little institute | ++ | Aspergillus fumigatus | In little institute | + |
| Candida albicans | In little institute | ++ | Aspergillus flavus | In little institute | + |
| Candida krusei | In little institute | + | Penicillium crysogenum | In little institute | + |
| Candida lipolytica | In little institute | +++ | Wood is mould | In little institute | - |
| The K yeast | In little institute | ++ | Aspergillus oryzae | In little institute | - |
| Italy's mould | In little institute | +++ | Aspergillus niger | In little institute | + |
| The pears phyllosticta | In little institute | ++ | The post spore is mould | In little institute | - |
| Ramularia panacicola | The Chinese Academy of Agricultural Sciences | ++ | Sturdy post is at a distance from spore | In little institute | - |
One, tetramycin is to the indoor biological activity determination test of Pyricularia oryzae
This experiment adopts growth rate method to carry out indoor bioactive mensuration; Clear and definite tetramycin is active to the inhibition of rice blast fungus; Confirm tetramycin to rice blast fungus EC50, EC70, EC90, for this product control rice blast provides high-tech data and foundation.
1, materials and methods
1.1 supply the examination material
Pyricularia oryzae, tetramycin, 4% kasugarnycin wetting powder
1.2 test method
1.2.1 pathogen is cultivated
Be inoculated in the medium after 2 rejuvenation of pathogen, cut the bacterium cake with the card punch of sterilizing, subsequent use.
1.2.2 supply the preparation of reagent agent
Tetramycin is configured to 5 gradients of 0.15,0.30,0.60,1.20,2.40 μ g/ml, and positive reference substance 4% kasugarnycin wetting powder is mixed with the concentration of 40.00 μ g/ml according to the field using dosage.
1.2.3 under aseptic condition, respectively with the sample and the reference substance of each concentration dose, add in the medium, fully mixing in the equivalent importing culture dish, is processed the pastille flat board of respective concentration.
Test is established the processing that adds the equivalent sterile water and is done blank.
6 repetitions are established in each processing.
1.2.4 under aseptic condition, the bacterium cake of the Pyricularia oryzae that has prepared is inoculated in the dull and stereotyped central authorities of pastille, mycelia faces up, and covers the ware lid, puts in 25 ℃ of incubators and cultivates.
1.2.5 investigation method: according to the growing state investigation pathogen mycelial growth situation of mycelia in the blank culture dish.Measure colony diameter with slide calliper rule, unit is a millimeter.Each bacterium colony is vertically measured diameter respectively once with right-angled intersection, gets its mean value.
1.3 mathematical statistics and analytical method
1.3.1 computational methods
According to investigation result, calculate each concentration of treatment according to formula (1) (2), the mycelial growth inhibition rate of Pyricularia oryzae, unit are percentage, result of calculation keeps behind the decimal point 2.
D＝D1-D2………………………………(1)
In the formula: D is that bacterium colony increases diameter
D1 is a colony diameter
D2 is a bacterium dish diameter
In the formula: I is a mycelial growth inhibition rate
D0 increases diameter for the blank mycelia
Dt increases diameter for the chemicals treatment mycelia
1.3.2 statistical analysis technique
Logarithm X value according to each concentration of tetramycin; With the probability value Y of handled to percentage mycelial inhibition; Obtain formula with mathematical statistics method and will remember virulence regression equation, obtain EC50, EC70, EC90, in order to estimate the inhibitory action of tetramycin rice blast pathogen.
Adopt the analysis of the duncan's method significance of difference.
2, result and conclusion
Experiment shows that the tetramycin variable concentrations is handled and all reached significance level.
Virulence regression equation: Y=4.0746+2.2304X, correlation coefficient 0.9836.
Calculate EC50=0.2600 μ g/ml, EC70=0.4467 μ g/ml, EC90=0.9761 μ g/ml according to equation, when promptly supplying 5769 times of reagent dilution agents, can reach 50% the inhibiting rate of Pyricularia oryzae; When supplying 3358 times of reagent dilution agents, the Pyricularia oryzae inhibiting rate is reached 70%; When supplying 1536 times of reagent dilution agents, the Pyricularia oryzae inhibiting rate is reached 90%.
Conclusion: tetramycin has higher biologically active to Pyricularia oryzae.When drug concentration was between 1.2-2.4 μ g/ml, tetramycin was superior to the inhibitory action of Pyricularia oryzae or approaches 4% kasugarnycin wetting powder, but effective ingredient consumption is starkly lower than positive control.
Two, tetramycin control rice blast application test example
1, experiment and demonstration 1
Test place: Donggang City, Dandong City, Liaoning Province chair circle town
Trial crops and kind: paddy rice
Controlling object: rice blast (Pyricularia oryzae Cav)
Test objective: tetramycin is carried out the field proving test of rice blast, and its safety is estimated.
Environmental condition: this plot is the rice transplanting field, April 15 2007 sowing date, May 17 field planting date, seeding row spacing 9 * 5cm, 50 kilograms of sealing fertilizers, 45 kilograms in urea.
Test medicine: tetramycin
Contrast medicament: 75% tricyclazole wettable powder
Consumption and processing numbering:
| Handle numbering | Medicament | Dispenser dosage | Formulation rate (active ingredient amount) |
| (1) | Tetramycin | 1000 times | 1.5mg/kg |
| (2) | Tetramycin | 1250 times | 1.2mg/kg |
| (3) | Tetramycin | 1500 times | 1.0mg/kg |
| (4) | Positive control 75% tricyclazole wettable powder | 2000 times | 375mg/kg |
| (5) | Blank | - | - |
Method for using: spray-on process: carry out the dispenser first time at rice blast morbidity eve or their early stage, dispenser is 2-3 time altogether, 7 days at interval.
Test period and spraying times:
Dispenser for the first time: on August 9th, 2007; Dispenser for the second time: on August 20th, 2007
Investigation method:, investigate twice respectively at August 10 day before yesterday after July 21, the secondary dispenser.
5 samplings in every sub-district, every 50 fringe, according to standard, the record incidence calculates disease index and control efficiency.
Stage division: 0 grade: anosis;
1 grade: every fringe loss is (morbidity of indivedual branch stalk) below 5%;
3 grades: every fringe loss 6-20% (other branch stalk morbidity about 1/3);
5 grades: every fringe loss 21-50% (fringe neck or main shaft morbidity, grain half is flat);
7 grades: every fringe loss 51-70% (fringe neck morbidity, most of grain is flat);
9 grades: every fringe loss 71-100% (morbidity of fringe neck, dead ears).
The drug effect computational methods:
In the formula: CK0, CK1 are respectively the forward and backward disease index of blank district dispenser; PT0, PT1 are respectively the forward and backward disease index of chemicals treatment district dispenser.
Result and conclusion:
(1), result
(2), conclusion
Adopt the Deng Kenshi duncan's new multiple range method that the result is carried out statistical analysis.
Show through variance analysis: test medicine maximum dose level control efficiency is higher than positive control sample, and the active ingredient formulation rate is starkly lower than positive control.Test medicine control rice blast is used 2-3 time in rice blast premorbid or initial stage dispenser continuously, and every mu of amount of liquid medicine is 50 liters, can effectively prevent rice blast, and in experimentation, paddy growth had no adverse effects, and does not see poisoning.
2, experiment and demonstration 2
Test place: experimental field, the agrotechnical center of Heilungkiang Fujin City
Trial crops and kind: paddy rice
Controlling object: rice blast (Pyricularia oryzae Cav)
Test objective: tetramycin is carried out the field proving test of rice blast, and its safety is estimated.
Environmental condition: the experimental field is ground, old rice field, and soil is white pulp grassy marshland, and clay fertilizer power is medium, and mu is executed fertile 25 kilograms of 40% soil prescription, 10 kilograms in urea.On April 15th, 2007 grew seedlings, rice transplanting on May 25, rice transplanting specification 30cm * 10cm---2-3 strain.
Test medicine: tetramycin
Contrast medicament: 25% hectogram missible oil
Consumption and processing numbering:
| Handle numbering | Medicament | Dispenser dosage | Formulation rate (active ingredient amount) |
| (1) | Tetramycin | 1000 times | 1.5mg/kg |
| (2) | Tetramycin | 1250 times | 1.2mg/kg |
| (3) | Tetramycin | 1500 times | 1.0mg/kg |
| (4) | Positive control 25% hectogram missible oil | 2000 times | 225mg/kg |
| (5) | Blank | - | - |
Method for using: spray-on process: foliar spray, do not handle 3 hydrargyrum oxydatum crudum liquid, repeat 4 times.
Test period and spraying times:
Leaf pest initial phase and July 30 days twice dispenser in July 21.
Investigation method: respectively at August 9, August 20, investigation on August 28 three times.
5 samplings in every sub-district, every 50 fringe, with back 7 days of last fertilising, according to standard, the record incidence calculated disease index and control efficiency before each fertilising.
Stage division: 0 grade: anosis;
1 grade: leaf spot lesion is less than 5, and length is less than 1cm;
3 grades: leaf spot lesion 6-10, part scab length is greater than 1cm;
5 grades: leaf spot lesion 11-25, the part scab is linked to be sheet, accounts for leaf area 10-25%;
7 grades: leaf spot lesion is more than 26, and scab is linked to be sheet, accounts for leaf area 26-50%;
9 grades: leaf spot lesion is linked to be sheet, accounts for leaf area more than 250%.
The drug effect computational methods:
In the formula: CK0, CK1 are respectively the forward and backward disease index of blank district dispenser; PT0, PT1 are respectively the forward and backward disease index of chemicals treatment district dispenser.
Result and conclusion:
1, result
2, conclusion
The survey showed that, and the contrast chemicals treatment shows tangible control efficiency to rice blast, and after the secondary dispenser 10 days is 82.66% to leaf seasonal febrile diseases preventive effect.
The test medicine tetramycin improves the rice blast preventive effect along with dispenser dosage increases.Experimental data is used Deng Kenshi duncan's new multiple range method statistic analysis result: 1000 times of dilution preventive effects of tetramycin effect is higher than the positive control medicament, and formulation rate is starkly lower than the positive control medicament.
Observe after the dispenser, it is normal that tetramycin is handled paddy growth, and no poisoning takes place.
From above result of the test, can draw the present invention and present compared with techniques, have following beneficial effect:
1, prove through indoor and field trial: 1000 times of dilution controls of tetramycin rice blast effect is superior to 4% kasugarnycin wetting powder, 75% tricyclazole wettable powder and 25% hectogram missible oil; And formulation rate is starkly lower than positive control; This is explanation just: use tetramycin not only can reduce agriculture production cost; Improve preventive effect, and can be used alternatingly, slow down the generation of resistance with other biopesticide;
2, tetramycin has inhibitory action to Gram-negative, positive bacteria, yeast and filamentous fungi (comprising disease fungus) etc.; And the apple canker control efficiency is reached more than 95%; Rice blast, alternaria leaf spot of apple and graw mold of tomato also there are good control efficiency, and have broad spectrum activity;
3, tetramycin is the produced simultaneously biological antibiotic that contains 4 kinds of compositions of a strain bacterium, belongs to environmentally friendly agricultural chemicals, and multiple antibiotic synergy, can reduce the use of other agricultural chemicals, has the certain significance to safeguarding ecotope;
4, tetramycin is a low toxicity through evidence.Acute toxicity LD50 is 4000mg/kg, all no abnormal reaction of skin and MMi, and Salmonella reversion test, SCE mutagenesis of CHL-1 cell and micronucleus test are all negative.The accumulation toxicity test check result shows the equal Non Apparent Abnormality of each organ, and cumulative coefficient is greater than 5.6, and more than explanation tetramycin toxicity is very low, and acute toxicity and salt and sodium bicarbonate are similar;
Embodiment
Through specific embodiment given below, can further understand the present invention.But they are not the restriction to protection domain of the present invention.
Embodiment 1: the tetramycin preparation of fermentation liquid
(1) seed fermentation medium: contain in per 100 milliliters: soybean cake powder 1.0g, starch 1.0g, glucose 0,7g; Peptone 0.5g, magnesium sulfate 0.03g, ammonium sulfate 1.0g, dipotassium hydrogen phosphate 0.02g; Calcium carbonate 0.7g, sodium chloride 0.1g, vegetable oil 0.2g, PH nature;
(2) submerged fermentation culture medium: contain in per 100 milliliters: glucose 2.5g, starch 1,5g, corn flour 1.0g; Soybean cake powder 2,0g, peptone (or corn steep liquor) 0.5g, ammonium sulfate 0.2g; Magnesium sulfate 0.03g, dipotassium hydrogen phosphate 0.02g, calcium carbonate 0.7g, sodium chloride 0.1g; Vegetable oil 0.2g, the normal propyl alcohol of culture volume 0.4% is added in the sterilization back, the PH nature.
(3) preparation of seed fermentation liquid: measure not streptomyces hygroscopicus Wuzhou subspecies (Streptomycesanygroscopicus subsp.Wuzhouensis n.subsp) bacterial strain by the inoculation of 5-20%; Cultivated 20-24 hour in the seed fermentation medium in 25-35 ℃; Ventilation: 1: 0.5-1 (V/V); Rotating speed: 150-300 rev/min, it is subsequent use to produce seed fermentation liquid;
(4) tetramycin preparation of fermentation liquid: measure above-mentioned seed fermentation liquid by the inoculation of 5-20% and carry out the submerged fermentation cultivation in 25-35 ℃; Ventilation: 0-12 hour early stage, 12-40 hour mid-term, later stage 40-42 hour: 1: 0.5-1 (V/V); Rotating speed: 150-300 rev/min; Inoculum concentration: 5-20% fermentation period: 64-72 hour, obtains the tetramycin zymotic fluid.
Embodiment 2-11: tetramycin control rice blast application example
Tetramycin control rice blast field control effectiveness test initial data statistical form (Donggang City, Dandong City, Liaoning Province chair circle town)
Tetramycin control rice blast field control effectiveness test initial data statistical form (experimental field, the agrotechnical center of Heilungkiang Fujin City)
Claims (1)
1. tetramycin preparation of fermentation liquid method that is used to prevent and treat rice blast is characterized in that it may further comprise the steps:
(1) seed fermentation medium: contain in per 100 milliliters: soybean cake powder 1.0g, starch 1.0g, glucose 0.7g, peptone 0.5g; Magnesium sulfate 0.03g, ammonium sulfate 1.0g, dipotassium hydrogen phosphate 0.02g, calcium carbonate 0.7g; Sodium chloride 0.1g, vegetable oil 0.2g, PH nature;
(2) submerged fermentation culture medium: contain in per 100 milliliters: glucose 2.5g, starch 1.5g, corn flour 1.0g, soybean cake powder 2.0g; Peptone or corn steep liquor 0.5g, ammonium sulfate 0.2g, magnesium sulfate 0.03g; Dipotassium hydrogen phosphate 0.02g, calcium carbonate 0.7g, sodium chloride 0.1g; Vegetable oil 0.2g, the normal propyl alcohol of culture volume 0.4% is added in the sterilization back, the PH nature;
(3) preparation of seed fermentation liquid: measure not streptomyces hygroscopicus Wuzhou subspecies (Streptomyces ahygroscopicus subsp.Wuzhouensis n.subsp) bacterial strain by the inoculation of 5-20%; Cultivated 20-24 hour in the seed fermentation medium in 25-35 ℃; Ventilation: 1: 0.5-1 (V/V); Rotating speed: 150-300 rev/min, it is subsequent use to produce seed fermentation liquid;
(4) tetramycin preparation of fermentation liquid: measure above-mentioned seed fermentation liquid by the inoculation of 5-20% and carry out the submerged fermentation cultivation in 25-35 ℃; 0-12 hour early stage, 12-40 hour mid-term, later stage 40-42 hour; Ventilation: 1: 0.5-1 (V/V), rotating speed: 150-300 rev/min, inoculum concentration: 5-20%; Fermentation period: 64-72 hour, obtain the tetramycin zymotic fluid.
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| CN102894000B (en) * | 2012-09-05 | 2015-01-07 | 青岛文创科技有限公司 | Tetramycin and tea saponin synergism pesticide composition |
| CN103314970A (en) * | 2013-06-19 | 2013-09-25 | 北京燕化永乐农药有限公司 | Bactericidal composition |
| CN103271094A (en) * | 2013-06-19 | 2013-09-04 | 北京燕化永乐农药有限公司 | Compound bactericide |
| CN104604895B (en) * | 2013-11-26 | 2017-05-10 | 辽宁微科生物工程股份有限公司 | Tetramycin compound and application |
| CN103636629A (en) * | 2013-11-28 | 2014-03-19 | 北京燕化永乐生物科技股份有限公司 | Polyoxin-containing bactericidal composition |
| CN103719113B (en) * | 2013-12-16 | 2015-11-18 | 骆毛喜 | A kind of aqueous pesticide solution and preparation method thereof and application |
| CN104430407B (en) * | 2014-12-18 | 2016-01-13 | 中国计量学院 | Tetramycin P is suppressing the application in Rhizoctonia solani and botrytis cinerea |
| CN104957153B (en) * | 2015-06-22 | 2018-06-19 | 广东中迅农科股份有限公司 | Composition pesticide containing tetramycin and fluorine azoles ring bacterium amine |
| CN107864983A (en) * | 2017-11-29 | 2018-04-03 | 孙铭岐 | A kind of biological pesticide for preventing and treating rice pest |
| CN109142589B (en) * | 2018-10-25 | 2022-03-01 | 湖南农业大学 | Method for determining tetramycin residual quantity in soil by using high performance liquid chromatograph |
| CN113321554A (en) * | 2021-06-29 | 2021-08-31 | 吉林省农业科学院 | Disease control agent and application thereof, rice seedbed conditioner and preparation and use methods thereof |
| CN115710144B (en) * | 2022-12-01 | 2024-04-30 | 贵州省烟草公司贵阳市公司 | Pesticide-fertilizer mixed combination for preventing and treating tobacco target spot disease and application thereof |
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| CN1172094A (en) * | 1996-07-25 | 1998-02-04 | 中国科学院沈阳应用生态研究所 | Multifunctional vegetable seedlings nutritious mother agent and its preparation method |
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