CN105175403A - Compound for inhibiting growth of bacterial wilt of kidney beans - Google Patents
Compound for inhibiting growth of bacterial wilt of kidney beans Download PDFInfo
- Publication number
- CN105175403A CN105175403A CN201510722683.6A CN201510722683A CN105175403A CN 105175403 A CN105175403 A CN 105175403A CN 201510722683 A CN201510722683 A CN 201510722683A CN 105175403 A CN105175403 A CN 105175403A
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- compound
- kidney bean
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- bacterial wilt
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
- C07D413/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings
- C07D413/04—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings directly linked by a ring-member-to-ring-member bond
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/72—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with nitrogen atoms and oxygen or sulfur atoms as ring hetero atoms
- A01N43/86—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with nitrogen atoms and oxygen or sulfur atoms as ring hetero atoms six-membered rings with one nitrogen atom and either one oxygen atom or one sulfur atom in positions 1,3
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Agronomy & Crop Science (AREA)
- Pest Control & Pesticides (AREA)
- Plant Pathology (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Dentistry (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Environmental Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
Abstract
The invention discloses a compound which has the inhibitory and bactericidal activity on bacterial wilt of kidney beans. The minimum inhibitory concentration (MIC) value of the bacterial wilt of the kidney beans is 4 micrograms per milliliter, and the minimal bactericidal concentration (MBC) value of the bacterial wilt of the kidney beans is 8 micrograms per milliliter.
Description
Technical field
The present invention relates to the novelty teabag of compound, illustrating the relation between pharmaceutical chemistry structure and the biological activity of bacterium, in particular, is explore compound to the antibacterial of Kidney bean bacterial wilting germ and fungicidal activity.
Background technology
Bean bacterial northern wilt is from Hedges1922 first since American South Dakota finds report, and become one of most important Micobial Disease of Kidney bean, the loss of some time almost reaches 100%.This disease mainly occurs in North America and Australia, in the U.S., mainly occurs in each state, northeast, then mainly occurs in its south and Victoria in Australia; In Europe, Turkey occurs serious, and all the other country damages are less.Bean bacterial northern wilt also occurs in south, Africa, causes fourth hairgrass wilt disease in Latin America.
According to Kennedy and Alcorn1980, the U.S. only Russia's carat river horse state in 1976, because bean bacterial northern wilt makes Kidney bean production loss reach 200,000 tons; When falling ill serious, Kidney bean the underproduction can reach 90%.Within 1975, in the Iowa of the U.S., 3 county's Late Cambrian wilting virus of kidney bean cause Soybean Brown Spot pinta, to nineteen eighty-three rapid spread to 16 counties in this state; Present Canada, USSR (Union of Soviet Socialist Republics) (particularly the Far East Area) all has generation.
It is reported, U.S. soybean production loss in 1975 reaches 13%.The susceptible Soybean Field in nineteen eighty-two Iowa reaches 84%, and wherein 1978 ~ 1981 years soybean yield loss reach 0 ~ 18.8%. and are respectively 7.8%(1978), 12.5%(1979), 4.3%(1981).According to records, some variety seeds susceptible gene reaches 30%.Wood etc. 1990 report, Queensland ,Australia finds a kind of new expression and pinto and mung bean wilt disease for 1984, and Infectikon rate reaches 5 ~ 90%.
The reported first in 1985 such as Chavarro, has found fourth hairgrass wilt disease in Latin America Colombia, and wilting virus of kidney bean infects fourth hairgrass and phaseolus vulgaris seeds reaches 88.8% and 52.5% respectively, rate of emergence degradation.
Wilting virus of kidney bean is bacillus pumilis, until slightly curved or wedge shape, and single or V, Y, paliform arrangement; On yeast extract paste glucose agar medium, bacterium colony is smooth, complete, lowly grandly not to glue, yellow, orange, pink or purple.
Wilting virus of kidney bean is mainly survived the winter on seed and sick stem.Germ survives with inner at the seed-coat of Kidney bean and soybean, and survival time can reach more than 20 years, can reach 24 years in laboratory conditions; According to records, when Kidney bean and beet crop rotation, pathogenic bacteria at least survives 2 winters in soil.Bean bacterial northern wilt is a typical vascular bundle diseases, mainly by seed dispersal.Seed cotyledons carries disease germs and passes to blade or directly enter vascular tissue, causes system to fall ill.Germ is infected mainly through wound again, and the incidence of wilting increases with the wound of stem and leaf tissue and increases.Under certain condition, the Meloidogyne incognita wound that provides cause of disease to invade.The favourable local propagation of irrigation water, the highlands as the U.S. only central and west regions and irrigation occurs general.Sand loam soil is heavier than clayed soil morbidity; Low temperature, rain and strong wind be conducive to disease occur and popular.
Summary of the invention
The present invention adopts In vitro Bactericidal Experiments, and research compound is to the biological activity of Kidney bean bacterial wilting germ.
Concrete technical scheme of the present invention is as follows:
Innovative point of the present invention finds that compound has good antibacterial and fungicidal activity to Kidney bean bacterial wilting germ, and measurement obtains its minimal inhibitory concentration MIC value and minimal bactericidal concentration MBC value, belongs to first public.
Described compound structure feature is shown below:
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 medium solid medium: get nutrient agar medium 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 Bechtop 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, microbial culture 24h, culture 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 trial-product.Bacterium adopts colony counting method to count, and by above bacterium liquid stroke-physiological saline solution redilution 100 times, gets 50 μ L, is evenly applied to and has been covered with in the plate of solid medium, and cultivate 24h, culture 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.
Calculation 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 minimum 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 different concns, 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, with visual inspection medicine minimum concentration Guan Zhongwu bacterial growth, person is the MIC of this trial drug, and each experiment in triplicate.
Recording minimal inhibitory concentration MIC value is 4 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 medium solid medium: get nutrient agar medium 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 Bechtop 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, microbial culture 24h, culture 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 trial-product.Bacterium adopts colony counting method to count, and by above bacterium liquid stroke-physiological saline solution redilution 100 times, gets 50 μ L, is evenly applied to and has been covered with in the plate of solid medium, and cultivate 24h, culture 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; Calculation 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 medium, continue to cultivate by under MIC culture condition, with the minimum concentration of complete kill bacteria for minimal bactericidal concentration (colony number 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 different concns, 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, and cultivate 24h in 28 DEG C after mixing, with visual inspection medicine minimum concentration Guan Zhongwu bacterial growth, person is the MIC of this trial drug.Meat soup in the above-mentioned each hole having no growth bacterium is got 10 μ L to be inoculated on nutrient agar plate, carries out mark, and cultivate 24h in 28 DEG C, to be still designated as the MBC of this medicine without drug level in the pipe of bacterial growth, each experiment in triplicate.
Recording minimal bactericidal concentration MBC value is 8 μ g/mL.
Claims (1)
1. the compound suppressing Kidney bean bacterial wilting germ to grow, is characterized in that:
(1) shown in following structural features formula:
(2) it can be used as inhibitor and the sterilant of Kidney bean bacterial wilting germ;
(3) it is 4 μ g/mL to the minimal inhibitory concentration MIC value of Kidney bean bacterial wilting germ;
(4) it is 8 μ g/mL to the minimal bactericidal concentration MBC value of Kidney bean bacterial wilting germ.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510722683.6A CN105175403A (en) | 2015-10-31 | 2015-10-31 | Compound for inhibiting growth of bacterial wilt of kidney beans |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510722683.6A CN105175403A (en) | 2015-10-31 | 2015-10-31 | Compound for inhibiting growth of bacterial wilt of kidney beans |
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| Publication Number | Publication Date |
|---|---|
| CN105175403A true CN105175403A (en) | 2015-12-23 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510722683.6A Pending CN105175403A (en) | 2015-10-31 | 2015-10-31 | Compound for inhibiting growth of bacterial wilt of kidney beans |
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Citations (2)
| 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 |
-
2015
- 2015-10-31 CN CN201510722683.6A patent/CN105175403A/en active Pending
Patent Citations (2)
| 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 |
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》 * |
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Application publication date: 20151223 |