CN105076163B - New application of the jamaicin as disinfectant use in agriculture - Google Patents
New application of the jamaicin as disinfectant use in agriculture Download PDFInfo
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Abstract
The invention belongs to pesticides application technical field, is related to the new application of jamaicin, particularly relates to new application of the jamaicin as disinfectant use in agriculture.New application of the jamaicin as disinfectant use in agriculture, the new application of described disinfectant use in agriculture are the purposes in the preventing and treating microbial plant disease of pathogenic.The present invention has found that jamaicin has good inhibiting effect to various plants pathogen by studying, and theoretical foundation is provided for jamaicin is developed into a kind of novel pesticide.
Description
Technical field
The invention belongs to pesticides application technical field, is related to the new application of jamaicin, particularly relates to jamaicin work
For the new application of disinfectant use in agriculture.
Background technology
Jamaicin is also known as berberine.It is a kind of common morphinane alkaloid, molecular formula C20H18NO4, its structure is as follows：
Jamaicin is present in many plants of the section ten of Berberidaceae etc. four category.M.-E. Xia Waliai and G. in 1826
Pei Ertan is obtained first from Xanthoxylonclava barks.Jamaicin is a kind of quartermary ammonium alkaloids.It can be separated out from ether
Yellow needle-like crystals；145 DEG C of fusing point；Water is dissolved in, is insoluble in benzene, ether and chloroform.It is mainly used in treating gastroenteritis, bacillary dysentery
The enteric infections such as disease, eye conjunctivitis, otitis media suppurative etc. are effective.Recently it has also been found that this product has blocking alpha-receptor, the anti-rhythm of the heart loses
Often effect.
Jamaicin has to hemolytic streptococcus, staphylococcus aureus, gonococcus and Freund, Shigella shigae etc.
Antibacterial action, and have enhancing white blood cell phagocytosis, also there is different degrees of inhibitory action to tubercle bacillus, plague bacillus, to big
The amoeba bacterium of mouse also has suppression effectiveness.Jamaicin has anti-curare to act in animals, and has peripheral decompression and solution
Heat effect.The hydrochloride (being commonly called as Halomine) of jamaicin is widely used in treatment gastroenteritis, bacillary dysentery etc., to lung knot
Core, scarlet fever, acute tonsillitis and respiratory tract infection also have certain curative effect.The traditional Chinese medical science is often with the coptis, golden cypress, barberry and ten
Big contribution etc. makees clearing heat and detoxicating medicine, and wherein principle active component is jamaicin.
At present, jamaicin is mainly used in field of medicaments, and the active material about jamaicin is in terms of Ag-fungicidal Activity
Research it is less.A variety of knurls of plant root rot are caused to obstruct infecting for spore it is reported that jamaicin can be resisted；Deng Yecheng etc. is ground
Study carefully discovery, Mahonia fortunei rhizome extract has good inhibiting effect to pears brown spot, and its median effective concentration is
1.798g/L。
The information for being disclosed in the background section is merely intended to understanding of the increase to the general background of the present invention, without answering
It has been the prior art well known to persons skilled in the art when being considered as recognizing or implying the information structure in any form.
The content of the invention
The purpose of the present invention is to expand the purposes of jamaicin, and theoretical base is provided as the purposes of disinfectant use in agriculture for jamaicin
Plinth.
Technical scheme provided by the invention is：
New application of the jamaicin as disinfectant use in agriculture, the new application of described disinfectant use in agriculture is in preventing and treating pathogenic
The purposes of microbial plant disease.
Preferably, described phytopathogen includes plant pathogenic fungi and plant pathogenetic bacteria.
Preferably, described plant pathogenic fungi is southern corn leaf blight, Curvularia lunata bacterium, Phytophthora nicotianae
Germ, citrus common green mold bacterium and tobacco sclerotium rolfsii.
Preferably, described plant pathogenetic bacteria is rice leaf spot bacteria, xanthomonas oryzae pv. oryzicola, Lee bacterium
Property perforation germ, walnut alternaria and citrus processing.
Compared with prior art, the present invention has the advantages that：
1. the jamaicin that the present invention uses extracting and developing from Mahonia fortunei obtains, for by narrow ten big work(of leaf
Labor exploitation is laid a good foundation into a kind of botanical fungicide, enriches the plant variety with bactericidal action.
2. the activity that inventor suppresses various plants pathogen to jamaicin has made intensive studies, experimental result is found
Jamaicin has good inhibiting effect to various plants pathogen, and the present invention carries for jamaicin is developed into a kind of novel pesticide
Theoretical foundation is supplied.
Embodiment
The embodiment of the present invention is described in detail with reference to embodiment, it is to be understood that the guarantor of the present invention
Shield scope is not limited by embodiment.
Explicitly indicated that unless otherwise other, otherwise in entire disclosure and claims, term " comprising " or its change
Change such as "comprising" or " including " etc. and will be understood to comprise stated element or part, and do not exclude other members
Part or other parts.
Inventor is had found that jamaicin has to various agricultural phytopathogen and preferably suppressed by substantial amounts of screening test
Effect, this is clear that from following biologicall test example.
Embodiment 1：Inhibitory action of the jamaicin to plant pathogenic fungi
With reference to Mu Liyi (1991) mycelial growth rate method, test test sample is given birth to different plant pathogenic fungi mycelia
The influence of long speed.
(1) for the activation of examination strain：It will be inoculated in for examination fungi in 9cm PSA flat board culture dishes, it is incubated in 28 DEG C
Cultivated in case, treat that bacterium colony length to the 2/3 of culture dish, is punched in colony edge with a diameter of 7mm sterilization punchers, diameter is made
It is standby for 7mm bacteria cake.
(2) preparation with malicious plating medium：Appropriate test sample is weighed, after being completely dissolved with methanol, with going out
Sample solution is made in the dilution of bacterium water, and the heat that pipette samples solution adds to quantification of 100mL is melted and is cooled to 50 DEG C or so of PSA
In hot culture medium, fully mix, pour into diameter 9cm culture dishes, the consistent band poison culture medium of thickness is made, fully solidifies standby
With.
(3) control is set：1. blank control (CK1)：Aqua sterilisa compares；2. solvent control (CK2)：Can dissolve for examination
Sample and to the solvent that has no significant effect of fungi growth as solvent control.
(4) processing step：Picking is inverted in the center with malicious plating medium, connect per ware for the bacteria cake of examination strain respectively
One bacteria cake, 3 repetitions are each handled, perform mark.Processing ware is positioned in 28 DEG C of constant incubators and cultivated, timing is seen
Examine colony growth situation.When the diameter of control group bacterium colony reaches more than the 2/3 of culture dish diameter, bacterium colony is measured with crossing method
Diameter, perform record.Using following equation, bacteriostasis rate of the active material to plant pathogenic fungi is calculated：
In the case where barberry alkali concn is 0.1g/L, jamaicin is tested to 16 kinds of phytopathies using above-mentioned mycelial growth rate method
The fungistatic effect of fungal pathogenses.As a result show, jamaicin has certain bacteriostasis (table 1) to 15 kinds of plant pathogenic fungis.Its
In, jamaicin shows as complete inhibition (inhibiting rate up to 100%) to southern corn leaf blight, to Curvularia lunata bacterium and cigarette
The inhibitory action of careless phytophthora germ is stronger, inhibiting rate more than 80%, also have to citrus common green mold bacterium and tobacco sclerotium rolfsii compared with
Good inhibitory action, inhibiting rate more than 60%, but jasmine phyllosticta leaf spot fungi, Mango are intended disk stey leaf spoting bacteria,
Kiwi berry rod spore is mould, the mould leaf spot fungi of tobacco rod spore, tobacco ash arrhizus bacteria, black spot of cabbage bacterium, Glorosprium musarum Cookeet Mass, rice rice
The inhibitory action of seasonal febrile diseases bacterium, withered germ of water-melon and citrus resin germ is general or poor, inhibiting rate between 6%~44%,
Almost there is no inhibitory action to Rhizoctonia solani Kuhn.Compared with clear water processing, solvent control used is to for trying pathogen without suppression
Make and use.
Bacteriostatic activity of the jamaicin of table 1 to plant pathogenic fungi
Embodiment 2：Inhibitory action of the jamaicin to bacterium
It is as follows with reference to the agar punching diffusion method in inhibition zone method, operating procedure：
(1) preparation of bacterium solution：To be rule activation for examination bacterium in NA culture mediums, be cultivated in 28 DEG C of constant incubators
48h, then be placed in 28 DEG C into NA Liquid Cultures with the sterilizing cultured bacterium of ring picking, vibrated in 120r/m shaken cultivation casees
24h is cultivated, it is standby.
(2) preparation with bacterium culture medium：Quantification of 100mL NA culture mediums heat is melted completely, treats that it is cooled to 45 DEG C
Left and right, draw in the cultured bacterial suspension injection culture mediums of 1mL, fully shake up, uniformly pour into a diameter of 9cm culture dishes
In, it is standby after solidification per ware about 25mL NA culture mediums.
(3) prepared by sample solution：The sample solution of fully dissolved is taken, the concentration for being configured to need with aqua sterilisa dilution is standby
With.
(4) control is set：(a) medicament control (CK1)：Positive control is used as using the medical streptomysins of 100mg/L；(b) clear water
Compare (CK2)：Aqua sterilisa compares；(c) solvent control (CK3)：Solvent to dissolve test sample is used as solvent control.
(5) processing step：A diameter of 5mm card punch is sterilized with alcolhol burner flame sterilization, after its cooling, carried disease germs
Punched on culture medium, beat 3-4 holes (triangular in shape or square) per ware, the culture medium in aperture is chosen away with the toothpick of sterilizing.Will
The sample solution and control solvent configured in orderly injection aperture, per the μ L of hole 45, carries out mark successively, each handles 3 times
Repeat.The ware handled well is placed in 28 DEG C of constant incubators after cultivating 48h, antibacterial circle diameter is measured using crossing method.
Concentration be 1g/L under, test jamaicin to 7 kinds of plant pathogenetic bacterias, 2 kinds of conventional plant disease biocontrol bacterias and
The bacteriostatic activity of this 2 kinds of common human pathogens of Escherichia coli and staphylococcus aureus.
Bacteriostatic activity of the jamaicin of table 2 to bacterium
As shown in table 2, jamaicin is respectively provided with certain bacteriostatic activity to 6 kinds of plant pathogenetic bacterias, wherein to the white leaf of rice
The fungistatic effect of rot bacterium is best, and antibacterial circle diameter reaches 32.00mm；Secondly it is bacillary for xanthomonas oryzae pv. oryzicola and Lee
Perforation germ, antibacterial circle diameter is respectively 14.17mm and 13.17mm；To walnut alternaria, staphylococcus aureus and citrus
The antibacterial circle diameter of ulcer bacteria is between 3-10mm；Growth to biocontrol microorganisms bacillus subtilis B47 and B96 does not almost have shadow
Ring, antibacterial circle diameter is respectively 1.50mm and 0.83mm；There is no fungistatic effect to Soft rot bacteria of Elephant taro and Escherichia coli.
Embodiment 3：Virulence of the jamaicin to xanthomonas oryzae pv. oryzicola
With reference to Du Lianxiang (2005) OD value method determination samples to the virulence for trying bacterium.
(1) prepared by strain：Actication of culture method is same as above, and the absorbance of pathogen is tested using UV detector, will
Bacterium solution is diluted to 1 × 109cfu/mL, standby.
(2) prepared by sample solution：The sample solution of fully dissolved is taken, the concentration for being configured to need with aqua sterilisa dilution, point
The drug containing Liquid Culture of various concentrations in 50mL liquid NA bottles the molten sample solvent of equivalent (control group contain), Jia Ru not be made
Base, it is standby.
(3) draw 0.5mL bacterium solutions to 50mL liquid NA cultivate in, it is 1 × 107cfu/mL bacteria suspensions that bacteria containing amount, which is made,.It is mixed
Close uniformly after 28 DEG C, 24h is cultivated under the conditions of 130r/m, it is to be measured.
(4) with the nutrient solution for not connecing bacterium containing same sample concentration do blank (control group molten sample solvent containing equivalent
Nutrient solution does blank) returned to zero, the OD600 values of processing are determined respectively.Bacteriostasis rate is calculated by following equation：
Using OD value methods, by pilot study, jamaicin is devised into five concentration by common ratio 1.68, respectively 1,
1.68th, 2.83,4.76 and 8mg/L；Comparison medicament is copper sulphate, also devises five concentration, respectively 2.4,2.76,3.174,
3.651 and 4.198mg/L.
Virulence of the jamaicin of table 3 to xanthomonas oryzae pv. oryzicola
As shown in table 3, jamaicin and copper sulphate are respectively 2.9008 Hes to the EC50 of xanthomonas oryzae pv. oryzicola
2.9929, compared with comparison medicament copper sulphate, jamaicin is stronger to the bacteriostatic activity of xanthomonas oryzae pv. oryzicola.
Embodiment 4：1% jamaicin microemulsion
Active compound jamaicin is dissolved in organic solvent and cosolvent, it is to be dissolved completely after, add emulsifying agent, stir,
It is allowed to form homogeneous mixed system, aqueous phase is then added dropwise into system, stirring forms transparent w/o type micro emulsion, then at 55 DEG C
Under the conditions of stirring in water bath, be allowed to form O/W type micro emulsions, that is, 1% jamaicin microemulsion be made.
Embodiment 5：Prevention effect of the 1% jamaicin microemulsion to bacterial leaf streak of rice
If 1% jamaicin microemulsion active ingredient 0.05g/L, 0.1g/L, 0.5g/L tri- is handled, can with 20% Yekuzuo
WP active ingredient 0.44g/L compares for medicament, using clear water processing as blank control, totally five processing, often handles 3 weights
It is multiple.Tillering stage rice is selected, pathogen is inoculated with to rice leaf using following two methods：
(1) stab inoculation：No. 5 insect needles two are taken, are fixed with rubber rubber head, distance controlling is in 0.5cm between two pins
Left and right.Bacterium solution (1.0x108cfu/mL) is dipped with pin, blade inoculation is pierced through in left and right sides by boundary of vein.Per repeated inoculation
15 leaves or 3 clumps of rice plants, after moisturizing culture 48h, dispenser preventing and treating.Scab length is measured after Routine Management 10d, 15d or 20d
Degree, prevention effect is calculated by following equation.
Prevention effect (%)=(1- processing scabs length/control scab length) × 100
(2) microtrauma mouth spray inoculation method：It is intended to Inoculated Rice plant and gathers discharge, with the smooth homogeneous iron in surface gently
Rice leaf is caught up with bat, is made to contact with each other between plant, blade, is rubbed, causes microtrauma mouth, the use of sprayer is then 1 by concentration
× 108cfu/mL bacterium solution is uniformly sprayed on the front and back of rice leaf, using not flow liquid as degree.6 clumps of rice are often repeated to plant
Plant, after moisturizing culture 48h, dispenser prevents and treats, and after Routine Management 15d, according to blank control incidence, is investigated according to disease scale
Method statistic data.The disease scale standard of bacterial leaf streak of rice is as follows：
0 grade：Blade face disease-free spot；
1 grade：Blade face only has the translucent water stain shape scab of dot, accounts for less than the 1% of leaf area；
3 grades：There is fragmentary short and fillet scab on blade face, accounts for the 1%~5% of leaf area；
5 grades：Blade face scab is more, accounts for the 6%~25% of leaf area；
7 grades：Blade face scab is closeer, accounts for the 26%~50% of leaf area；
9 grades：Blade face scab is distributed in more than 50%, and blade becomes tenne, crimps, be withered.
Drug effect computational methods：The data of pilot survey share disease index (hereinafter referred to as disease refers to) and prevention effect is (following
Abbreviation preventive effect) 2 kinds, result of the test is calculated according to the following formula；
Prevention effect of the needle point method inoculation test Berberine of table 4 to bacterial leaf streak of rice
As shown in table 4, the jamaicin of various concentrations is different to the prevention effect of bacterial leaf streak of rice, in three concentration
In, preventive effect highest when concentration is 0.5g/L, prevention effect reaches 94.06% during 10d after medicine, compared with Yekuzuo, difference
The not up to level of signifiance, but 20d drug effects decrease after medicine, prevention effect 89.04%, hence it is evident that less than Yekuzuo in concentration
For 0.44g/L when prevention effect.After medicine during 10d, the prevention effect difference of three concentration processing of jamaicin is little, does not reach
To the level of signifiance.After medicine during 20d, under 5% flat standard, the prevention effect between three concentration processing has reached notable water
It is flat.
Prevention effect of the 1% jamaicin microemulsion to bacterial leaf streak of rice in the spray-on process inoculation test of table 5
As shown in table 5, the prevention effect of spray inoculation method not as the prevention effect of needle point method it is good, but between three concentration
Prevention effect is similar with the sequence of needle point method prevention effect, is all that the higher preventive effect of concentration is better.Its Berberine 0.5g/L
Difference between 0.1g/L processing is not notable and suitable with the prevention effect of 0.44g/L Yekuzuos.
Embodiment 6：Jamaicin is to the lasting period of bacterial leaf streak of rice, protective effect and therapeutic action
The jamaicin microemulsion of table 6 1% is to the lasting period of bacterial leaf streak of rice, prevention and therapeutic effect
As shown in table 6, after 1% jamaicin microemulsion spraying treatment 21d, 17d, 14d, 11d, 7d, 3d and on the day of dispenser, adopt
It is inoculated with needle point method, and is sprayed after inoculation after 3d, 7d in 9 processing such as jamaicin, meets the place of bacterium on the day of spraying jamaicin
Manage best to the prevention effect of bacterial leaf streak of rice, preventive effect is up to 74.84%, but with spraying after jamaicin 3d, 7d, 11d again
The preventive effect of bacterium is connect compared to not reaching the difference pole level of signifiance；After dispenser 21d connect again bacterium prevention effect it is worst, preventive effect only has
34.74%.Although illustrating that jamaicin reduces to the prevention effect of bacterial leaf streak of rice with the extension of administration time,
Lasting period is longer；The prevention effect (63.67%) of dispenser is although be followed by the preventive effect of bacterium less than dispenser 7d after inoculation 3d
(70.77%) significance level of difference is not reached, but；Connect 7d dispensers after bacterium connect again with 14d after medicine bacterium prevention effect it is suitable, respectively
For 53.45% and 53.44%, this explanation jamaicin has preferable protecting effect and therapeutic effect to bacterial leaf streak of rice,
Protecting effect is better than therapeutic effect.
In summary, jamaicin has good inhibiting effect to various agricultural phytopathogen, especially thin to rice
The prevention effect of bacterium property Population of Xanthomonas Oryzae Pv is more notable, and one is provided for jamaicin is developed into a kind of new botanical fungicide
The new thinking of kind.
The description of the foregoing specific illustrative embodiment to the present invention is to illustrate and the purpose of illustration.These descriptions
It is not wishing to limit the invention to disclosed precise forms, and it will be apparent that according to above-mentioned teaching, can be much changed
And change.The purpose of selecting and describing the exemplary embodiment is that explain that the certain principles of the present invention and its reality should
With so that those skilled in the art can realize and utilize the present invention a variety of exemplaries and
Various chooses and changes.The scope of the present invention is intended to be limited by claims and its equivalents.
Claims (1)
1. new application of the jamaicin as disinfectant use in agriculture, it is characterised in that：The new application of described disinfectant use in agriculture is anti-
Control the purposes of the microbial plant disease of pathogenic；Described phytopathogen is plant pathogenetic bacteria；Described phytopathy
Indigenous bacteria is rice leaf spot bacteria, xanthomonas oryzae pv. oryzicola, Lee's Bacterial septicaemia syndrome bacterium, walnut alternaria and citrus
Ulcer bacteria.
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| CN109757470A (en) * | 2019-01-24 | 2019-05-17 | 北京联合大学 | Jamaicin corrigent, the botanical pesticide raw material containing the corrigent and preparation process |
| CN111184010B (en) * | 2020-01-19 | 2021-09-10 | 广西大学 | Pesticide composition containing berberine |
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| CN100581367C (en) * | 2007-05-11 | 2010-01-20 | 山东省农业科学院植物保护研究所 | Compound against plant virus |
| CN101731288B (en) * | 2008-11-20 | 2011-11-23 | 兰州世创生物科技有限公司 | Natural biological composition for preventing and treating agricultural pests and preparation method thereof |
| CN101433221B (en) * | 2008-12-09 | 2012-11-21 | 华中科技大学 | Chinese medicine bactericidal agent for preventing and treating soft rot of konjak |
| CN101695301B (en) * | 2009-09-29 | 2013-06-05 | 北京联合大学生物化学工程学院 | Micro-emulsion for killing Monilinia fructicola and preparation method thereof |
| CN101961019B (en) * | 2010-10-29 | 2013-02-13 | 湖北太极生化开发有限公司 | Plant source bactericide and application thereof |
| CN102349520B (en) * | 2011-08-25 | 2013-09-04 | 华南农业大学 | Application of berberine as herbicide |
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