CN105211078A - A kind of compound suppressing larch pin spoting disease bacteria growing - Google Patents
A kind of compound suppressing larch pin spoting disease bacteria growing Download PDFInfo
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- CN105211078A CN105211078A CN201510723794.9A CN201510723794A CN105211078A CN 105211078 A CN105211078 A CN 105211078A CN 201510723794 A CN201510723794 A CN 201510723794A CN 105211078 A CN105211078 A CN 105211078A
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- larch
- spoting
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- disease
- bacterium
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- 241000894006 Bacteria Species 0.000 title claims abstract description 40
- 241000218652 Larix Species 0.000 title claims abstract description 22
- 235000005590 Larix decidua Nutrition 0.000 title claims abstract description 22
- 201000010099 disease Diseases 0.000 title claims abstract description 21
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 title claims abstract description 21
- 150000001875 compounds Chemical class 0.000 title claims abstract description 11
- 230000000844 anti-bacterial effect Effects 0.000 claims abstract description 17
- 230000002401 inhibitory effect Effects 0.000 claims abstract description 9
- 239000003899 bactericide agent Substances 0.000 claims 1
- 239000003112 inhibitor Substances 0.000 claims 1
- 239000007788 liquid Substances 0.000 description 31
- 230000001580 bacterial effect Effects 0.000 description 13
- 235000015097 nutrients Nutrition 0.000 description 12
- 239000003814 drug Substances 0.000 description 9
- 239000002504 physiological saline solution Substances 0.000 description 8
- 238000002360 preparation method Methods 0.000 description 8
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- 229940079593 drug Drugs 0.000 description 5
- 239000001963 growth medium Substances 0.000 description 5
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- 239000012895 dilution Substances 0.000 description 4
- 238000010790 dilution Methods 0.000 description 4
- 239000012153 distilled water Substances 0.000 description 4
- 239000012467 final product Substances 0.000 description 4
- 230000001954 sterilising effect Effects 0.000 description 4
- 238000004659 sterilization and disinfection Methods 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 3
- 244000052769 pathogen Species 0.000 description 3
- 230000001717 pathogenic effect Effects 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 235000011613 Pinus brutia Nutrition 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 238000003113 dilution method Methods 0.000 description 2
- 239000012456 homogeneous solution Substances 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 238000005142 microbroth dilution method Methods 0.000 description 2
- 239000013642 negative control Substances 0.000 description 2
- 239000013641 positive control Substances 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 208000035473 Communicable disease Diseases 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241001006839 Globisporangium ultimum Species 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 241000544657 Larix gmelinii Species 0.000 description 1
- 241001139304 Larix gmelinii var. olgensis Species 0.000 description 1
- 241000411840 Larix gmelinii var. principis-rupprechtii Species 0.000 description 1
- 241000218641 Pinaceae Species 0.000 description 1
- 235000008331 Pinus X rigitaeda Nutrition 0.000 description 1
- 241000018646 Pinus brutia Species 0.000 description 1
- 244000007853 Sarothamnus scoparius Species 0.000 description 1
- 241000270708 Testudinidae Species 0.000 description 1
- 241000607479 Yersinia pestis Species 0.000 description 1
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- 230000000694 effects Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
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- 229920005989 resin Polymers 0.000 description 1
- 235000014347 soups Nutrition 0.000 description 1
- 239000002023 wood Substances 0.000 description 1
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- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The present invention finds to have compound that is antibacterial and bactericidal activity to larch pin spoting disease bacterium.The minimal inhibitory concentration MIC value of its larch pin spoting disease bacterium is 0.5 μ g/mL, and minimal bactericidal concentration MBC value is 1 μ g/mL.
Description
Technical field
The present invention relates to the novelty teabag of compound, illustrate the relation between pharmaceutical chemistry structure and the biologically active of bacterium, in particular, is explore compound to the antibacterial of larch pin spoting disease bacterium and bactericidal activity.
Background technology
The larch subfamily of Larch in Pinaceae, being the main forest composition seeds of Alpine coniferous forests in China northeast, forest zone, the Inner Mongol and North China, southwest, is one of large needle yielding timber plantationsion in the Northeast main three.The germ plasm resource of this genus is enriched, and having a very wide distribution, throughout the alpine region, subtropics in the Northern Hemisphere and the flat area of cool temperate zone, be originally mainly distributed in northeast in China, is the main group of Xing'anling mountains height above sea level 300 ~ 1200 meter band.Present distribution oneself expand, in Inner Mongolia Autonomous Region, Liaoning Province, Jilin Province, Heilongjiang Province, Shandong Province, Shaanxi Province, Gansu Province, Qinghai Province, Ningxia Hui Autonomous Region all have it to distribute.Larch is the most cold-resistant in coniferous species, and vertical distribution reaches the most upper limit of forest restoration.Dahurian larch, larix olgensis, Larix principis-rupprechtii, larch-tree and Korea's larch etc. are had in the northern area of China natural distributed and tame main larch.It has strong adaptability, wide accommodation, fast growth, and grow into forest early, the good characteristics such as disease and insect resistance, its strength of wood is high, and corrosion resistance is strong, industrially has been widely used, and can as excellent paper making raw material.Because it has above good characteristic, it can be used as large area reproducting tree species.
Larch pin spoting disease is one of important disease of larch.Tortoise wells in 1938 and aboveground this is sick in Hokkaido, Japan Late Cambrian, its pathogen is determined in field, nineteen fifty pool, extensively to infect cause great infringement to the sixties in Japan.Larch pin spoting disease is in the initial stage seventies since China starts, and once between the more than ten years, namely oneself spread all over the vast forest zone of three provinces in the northeast of China, is that harm is serious, spreads dangerous disease rapidly.
This disease causes harm the young sprout then of 1 ~ 35 year raw LARCH PLANTATION, and the young growth morbidity of life in 6 ~ 15 years is heavier and general.Originally move back green in the not lignified tender stem of young sprout or caulom portion, become crineous, black from light brown, micro-contraction attenuates.Often have resin to overflow, upper bend becomes hook-shaped, and leaf is withered, and major part comes off, only a residual parallel needles leaf at top.Pine needle produces the conidiospore device that black point is pathogen.Therefore, branch more than site of pathological change is withered, and seedling is become without top seedling.In the Second Year bark of spring on the sick tip stitches, the large stain of generation and pore are dothithecium and the conidiospore device of pathogen.
This disease not only causes larch young sprout withered, and more seriously, owing to being injured year after year, make tree crown bald, be deformed into broom shape, the diameter of a cross-section of a tree trunk 1.3 meters above the ground, the volume of timber, the height of tree decline year by year, and cause forest land not grow into forest, forest can not become a useful person.Given this disease is a kind of dangerous infectious disease, very harmful, country is formal is already classified as quarantine object, and on August 12nd, 2004, the State Administration of Forestry does in coinage [2004] No. 59 file distribution No. 4 " bulletin " Forest Plant Quarantine Pest, larch pin spoting disease is still among them, and absolutely proves the serious harm of this disease.
Summary of the invention
The present invention adopts In vitro Bactericidal Experiments, and research compound is to the biologically active of larch pin spoting disease bacterium.
Concrete technical scheme of the present invention is as follows:
Innovative point of the present invention finds that compound has good antibacterial and bactericidal activity to larch pin spoting disease bacterium, and measurement obtains its minimal inhibitory concentration MIC value and minimal bactericidal concentration MBC value, belongs to first public.
Described compound structure feature following graphic shown in:
Embodiment
Below in conjunction with concrete embodiment, in further detail the present invention is described.Embodiment below should be understood and be only not used in the restriction scope of the invention for illustration of the present invention.
embodiment 1
Measure minimal inhibitory concentration MIC value.
(1) preparation of nutrient broth: get nutrient broth 30g and add 1000mL distilled water and get final product.Use front 121 DEG C of high pressure steam sterilization 20min stand-by.
(2) preparation of nutrient agar solid culture medium: get nutrient agar 45g and add 1000mL distilled water and get final product.Use front 121 DEG C of high pressure steam sterilization 20min stand-by.
(3) cultivation of bacterial strain: operate on superclean bench and carry out.Drawing sterilized liquid nutrient medium l0mL, be placed in sterilized test tube, then with connecing collarium picking bacterium colony, being added in liquid nutrient medium, be put in incubator and cultivate, bacterial culture 24h, cultivation temperature is 28 DEG C.
(4) preparation of bacterium liquid and counting: by the bacterium liquid after cultivation, Bian 10 times of dilution method liquid nutrient mediums dilute, and with blood counting chamber preliminary observation counting on microscope, then bacterium liquid liquid nutrient medium is diluted, as the bacterium liquid added in test sample.Bacterium adopts colony counting method to count, and above bacterium liquid stroke-physiological saline solution is diluted 100 times again, gets 50 μ L, be evenly applied to and be covered with in the plate of solid culture medium, and cultivate 24h, cultivation temperature is 28 DEG C.After cultivation, single bacterial growth alive forms a bacterium colony, statistics colony count, can calculate in sample containing bacterium number.
Computing formula is: bacterial concentration=n × 20 × 100cfu/mL
(5) preparation of drug solution: Weigh Compound, adds sterile saline, shakes hook, obtains homogeneous solution, for subsequent use.Deposit in 4 DEG C of Refrigerator stores stand-by.
(6) mensuration of minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC): adopt micro-broth dilution method minimal inhibitory concentration MIC(MinimalInhibitoryConcentration).MIC is minimal inhibitory concentration, namely after medicine and the effect of certain density bacterium liquid, can suppress the least concentration of visible bacteria growing.
Adopt doubling dilution by liquid stroke-physiological saline solution by liquid dilution series concentration, 0.5,1,2,4,8,16,32,64,128 and 256 μ g/mL, 1 ~ 10 row on 96 orifice plates, every hole adds liquid and the 100 μ L bacterium liquid of 100 μ L variable concentrations, makes final bacterial concentration be 1 ~ 5 × 10
5cfu/mL, 11st row adds bacterium liquid as positive control using stroke-physiological saline solution, 12nd row is not to add the stroke-physiological saline solution of bacterium liquid for negative control, 24h is cultivated in 28 DEG C after mixing, to visually observe the MIC that medicine least concentration Guan Zhongwu bacterial growth person is this trial drug, each experiment in triplicate.
Recording minimal inhibitory concentration MIC value is 0.5 μ g/mL.
embodiment 2
Measure minimal bactericidal concentration MBC value.
(1) preparation of nutrient broth: get nutrient broth 30g and add 1000mL distilled water and get final product.Use front 121 DEG C of high pressure steam sterilization 20min stand-by.
(2) preparation of nutrient agar solid culture medium: get nutrient agar 45g and add 1000mL distilled water and get final product.Use front 121 DEG C of high pressure steam sterilization 20min stand-by.
(3) cultivation of bacterial strain: operate on superclean bench and carry out.Drawing sterilized liquid nutrient medium l0mL, be placed in sterilized test tube, then with connecing collarium picking bacterium colony, being added in liquid nutrient medium, be put in incubator and cultivate, bacterial culture 24h, cultivation temperature is 28 DEG C.
(4) preparation of bacterium liquid and counting: by the bacterium liquid after cultivation, Bian 10 times of dilution method liquid nutrient mediums dilute, and with blood counting chamber preliminary observation counting on microscope, then bacterium liquid liquid nutrient medium is diluted, as the bacterium liquid added in test sample.Bacterium adopts colony counting method to count, and above bacterium liquid stroke-physiological saline solution is diluted 100 times again, gets 50 μ L, be evenly applied to and be covered with in the plate of solid culture medium, and cultivate 24h, cultivation temperature is 28 DEG C.After cultivation, single bacterial growth alive forms a bacterium colony, statistics colony count, can calculate in sample containing bacterium number; Computing formula is: bacterial concentration=n × 20 × 100cfu/mL.
(5) preparation of drug solution: Weigh Compound, adds sterile saline, shakes hook, obtains homogeneous solution, for subsequent use.Deposit in 4 DEG C of Refrigerator stores stand-by.
(6) mensuration of minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC): adopt micro-broth dilution method minimal bactericidal concentration MBC(MinimalBactericidalConcentration).On MIC basis, draw 10 μ L solution from every pipe, put on solid culture medium, continue to cultivate by under MIC condition of culture, with the least concentration of complete kill bacteria for minimal bactericidal concentration (clump count is less than or equal to 5).
Adopt doubling dilution by liquid stroke-physiological saline solution by liquid dilution series concentration, 0.5,1,2,4,8,16,32,64,128 and 256 μ g/mL, 1 ~ 10 row on 96 orifice plates, every hole adds liquid and the 100 μ L bacterium liquid of 100 μ L variable concentrations, makes final bacterial concentration be 1 ~ 5 × 10
5cfu/mL, 11st row adds bacterium liquid as positive control using stroke-physiological saline solution, 12nd row, not add the stroke-physiological saline solution of bacterium liquid for negative control, cultivates 24h, to visually observe the MIC that medicine least concentration Guan Zhongwu bacterial growth person is this trial drug in 28 DEG C after mixing.Meat soup in the above-mentioned each hole having no growth bacterium is got 10 μ L to be inoculated on nutrient agar panel, carries out mark, and cultivate 24h in 28 DEG C, to be still designated as the MBC of this medicine without drug concentration in the pipe of bacterial growth, each experiment in triplicate.
Recording minimal bactericidal concentration MBC value is 1 μ g/mL.
Claims (1)
1. suppress a compound for larch pin spoting disease bacteria growing, it is characterized in that:
(1) following structural features graphic shown in:
(2) it can be used as inhibitor and the bactericide of larch pin spoting disease bacterium;
(3) it is 0.5 μ g/mL to the minimal inhibitory concentration MIC value of larch pin spoting disease bacterium;
(4) it is 1 μ g/mL to the minimal bactericidal concentration MBC value of larch pin spoting disease bacterium.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510723794.9A CN105211078A (en) | 2015-11-01 | 2015-11-01 | A kind of compound suppressing larch pin spoting disease bacteria growing |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510723794.9A CN105211078A (en) | 2015-11-01 | 2015-11-01 | A kind of compound suppressing larch pin spoting disease bacteria growing |
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| CN105211078A true CN105211078A (en) | 2016-01-06 |
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| CN201510723794.9A Pending CN105211078A (en) | 2015-11-01 | 2015-11-01 | A kind of compound suppressing larch pin spoting disease bacteria growing |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107022494A (en) * | 2017-04-05 | 2017-08-08 | 北京环氧环保科技发展有限公司 | One plant of pine the born of the same parents bacterium BN h1 of color two and biological bacteria degradation agent and its preparation method and application |
| CN114467939A (en) * | 2022-02-18 | 2022-05-13 | 东北林业大学 | Application of quinic acid as bacteriostatic agent in inhibiting growth of larch gluconobacter |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0360417A2 (en) * | 1988-08-24 | 1990-03-28 | Schering Agrochemicals Limited | Derivatives of 4-fluoroanthranilic acid and their use as fungicides |
| CN1344259A (en) * | 1999-02-16 | 2002-04-10 | 巴斯福股份公司 | Processes and intermediates for prepn. of 1,3-diazin-b 6-ones and uracils |
| CN1562080A (en) * | 2004-04-02 | 2005-01-12 | 吉林大学 | Broad spectrum bacteriostat and sterilizing of silicate based angstrom silver ion nano porous composite mateiral |
| CN104788428A (en) * | 2015-04-24 | 2015-07-22 | 南京农业大学 | Pyridazinone dipyrrolidone derivative, as well as preparation method and application thereof |
-
2015
- 2015-11-01 CN CN201510723794.9A patent/CN105211078A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0360417A2 (en) * | 1988-08-24 | 1990-03-28 | Schering Agrochemicals Limited | Derivatives of 4-fluoroanthranilic acid and their use as fungicides |
| CN1344259A (en) * | 1999-02-16 | 2002-04-10 | 巴斯福股份公司 | Processes and intermediates for prepn. of 1,3-diazin-b 6-ones and uracils |
| CN1562080A (en) * | 2004-04-02 | 2005-01-12 | 吉林大学 | Broad spectrum bacteriostat and sterilizing of silicate based angstrom silver ion nano porous composite mateiral |
| CN104788428A (en) * | 2015-04-24 | 2015-07-22 | 南京农业大学 | Pyridazinone dipyrrolidone derivative, as well as preparation method and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| KIM, MIN CHEOL ET.AL: "Salinazinones A and B: pyrrolidinyl-oxazinones from solar saltern-derived Streptomyces sp. KMF-004", 《ORGANIC LETTERS》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107022494A (en) * | 2017-04-05 | 2017-08-08 | 北京环氧环保科技发展有限公司 | One plant of pine the born of the same parents bacterium BN h1 of color two and biological bacteria degradation agent and its preparation method and application |
| CN114467939A (en) * | 2022-02-18 | 2022-05-13 | 东北林业大学 | Application of quinic acid as bacteriostatic agent in inhibiting growth of larch gluconobacter |
| CN114467939B (en) * | 2022-02-18 | 2023-03-28 | 东北林业大学 | Application of quinic acid as bacteriostatic agent in inhibiting growth of larch grape-seat bacteria |
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