CN100586286C - Plant antiviral agent, preparation method and application thereof - Google Patents
Plant antiviral agent, preparation method and application thereof Download PDFInfo
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- CN100586286C CN100586286C CN200710090014A CN200710090014A CN100586286C CN 100586286 C CN100586286 C CN 100586286C CN 200710090014 A CN200710090014 A CN 200710090014A CN 200710090014 A CN200710090014 A CN 200710090014A CN 100586286 C CN100586286 C CN 100586286C
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Abstract
The invention discloses a plant active ingredient of Cynanchum Komarovii extract with 7-defluvium unguium oxyl tylophorine as effective component and accessory ingredient of plant source antiviral agent preferably water, emulsifiable concentrate and wettable powder; preparation method and application against plant virus.Through indoor farm testing, the antiviral preparation can be used for preventing and treating pathology caused by single virus infecting different host or two or more viruses mixed infecting the same host or homogeneous host, especially plant pathology caused by 5 virus distributing world widely with severe hazard. The preventing and treating effect can reach up to 40-50%. The antiviral preparation has ideal effect in preventing and treating multi-viruses mixed infected tobacco. The inventive antiviral agent has no toxicity to human and livestock and high safety and performance.
Description
Technical field
The present invention relates to a kind of plant source disease-resistant toxic agent, wherein containing with the 7-demethoxytylophorine is the Cynanchun komarovii extract of active ingredient, and optionally contains auxiliary agent.Antivirotic of the present invention has good antiviral activity, is natural, efficient, safe plant source disease-resistant poison pesticidal preparations.
Background technology
Because the intrinsic defective of chemicals, the BIOLOGICAL CONTROL technology of aspects such as desinsection, sterilization and weeding is developed apace in recent years, and the disease pest and weed in many agricultural productions has all obtained control effectively.Yet because the particularity of structure of plant virus and physiology, biochemical aspect, the human at present preparation that does not also find efficient, safe anti-plant viral disease, thereby plant virus is also just more and more big to the harm of agricultural and the loss that caused, therefore at present to the very urgent (Zhu Shuifang of needs of the preparation of efficient, safe anti-plant viral disease, world agriculture, 1989,1,40-42; SunsunKubo plants the disease newspaper day, and 1990,65,481-487).
According to＜Inner Mongol flora〉record: " Cynanchun komarovii belongs to the square-bottomed bamboo basket wild poisonous herbaceous plant of section goose that rubs, formal name used at school Cynanchum komarovil AL.lijinski, another name Cynanchum Komarrivii AI Iijiniski, Lu Xin grass.Be distributed widely in unconcerned area, NORTHWEST CHINA drought sandy wasteland, Inner Mongol resource You Feng.The blue or green young stem and leaf livestock of this grass does not eat, and the autumn and winter can be made hay, and seed oil content 30% can be done iundustrial oil.Are good nectariferous plants six flowering stage in July.The Ningxia peasant comes Ih Ju League, Inner Mongol harvesting autumn grass, spreads paddy field after the chopping and can reduce damage by disease and insect and obtain volume increase.Use for reference experience among the people, in conjunction with experimental basis, the patent of invention ZL95118353.2 of Yao Yucheng etc. discloses the compound plant insecticide by Cynanchun komarovii and other plant preparation.
The common patent of invention ZL00106234.4 of Nankai University and Inner Mongol University of Technology discloses that main active is 7-demethoxytylophorine (Antofine) in the alcohol extract of Cynanchun komarovii, and main giving birth to according to exsomatizing surveyed data and determined that it is applied to the particularly control of tobacco mosaic virus disease (TMV) of the viroses of plant.After this, the application for a patent for invention CN200410060104.8 of Inst. of Pesticides, Xibei Univ. of Agricultural ﹠. Forestry Science ﹠. Technolo discloses the plant herbicide that contains Cynanchun komarovii extract; The application for a patent for invention CN200510096018.7 of Northwest University discloses a plant species environment-friendly pesticide fishing medicine, wherein contains the extract of kaladana and Cynanchun komarovii.Described weed killer herbicide and fishing medicine are main active ingredient with the 7-demethoxytylophorine all.
Tobacco mosaic virus disease is that tobacco cultivation district, the whole world takes place generally to endanger a serious big class disease, and the world has found and reported that the virus that infects tobacco reaches 47 kinds at present, and China has 17 kinds.It is wide wherein to distribute, endanger serious have three kinds, comprise cucumber mosaic virus (CMV), tobacco mosaic virus (TMV) and the mix dip-dye of marmor upsilon (PVY): in the popular time of disease to tobacco, the grave illness field underproduction 20～50%, the situation of the summer cigarette underproduction more than 70%.This has proved that a kind of virus can infect the host plant of multiple class, and endanger serious is that a certain host plant is simultaneously by the mixed infection of multiple virus, due to illness poison can only rely on host's biology finish it self duplicate, so in rapidly a large amount of virus of proliferation, must cause the heavy losses of host crop aspect quality and output, and this just virus host in plant structure, physiology, biochemical aspect the concrete manifestation of particularity.Above-mentioned three kinds of viruses are the widely distributed pattern samples that endanger three serious Tobamovirus of the same name again of world wide.No matter they are to infect separately or mixed infection, all are to make relevant host plant that the root of disease and multiple diseases take place, and serious situation is multiple viral mixed infection one plant species, can cause tobacco, melon, banana, pulse family, Solanaceae, the Chenopodiaceae various vegetables, flower ornamental nursery stock, fruit tree crop etc., heavy losses aspect quality and output, even reach destructive degree.The mankind press for efficiently, safe and eco-friendly new antiviral drug, but do not occur the Pesticidal products of plant source disease-resistant toxin preparation so far both at home and abroad.
The inventor is for being the Cynanchun komarovii extract of active ingredient with the 7-demethoxytylophorine, or make the preparation classification of the former medicine of total alkali and Antiphytoviral performance through separation and carried out the system research that comprises that indoor external test is tested, find that the Cynanchun komarovii extract is in this preparation system, preferred concentration interval below 100ppm, other disease of unitary class that various plants virus and described virus are caused on different plants or the disease of plurality of classes have good control efficiency, thereby implement and expanded the range of application of Cynanchun komarovii extract.
Summary of the invention
The purpose of this invention is to provide a kind of plant source disease-resistant toxic agent, it is the Cynanchun komarovii extract of active ingredient that described antivirotic contains with the 7-demethoxytylophorine, and optionally contains auxiliary agent.
The invention provides a kind of plant source disease-resistant toxic agent, wherein containing with the 7-demethoxytylophorine is that the former medicine of Cynanchun komarovii extract of active ingredient is an active component, gross weight in antivirotic, plant source disease-resistant toxic agent of the present invention contains the former medicine of 0.1-75%, and the auxiliary agent of 99.9-25%, the content of 7-demethoxytylophorine is about 0.03-5% (weight) in the wherein said former medicine.
Antivirotic of the present invention has good antiviral activity, is natural, efficient, safe plant source disease-resistant toxic agent.
Above use term " the former medicine of Cynanchun komarovii extract " to be meant with the Cynanchun komarovii to be the extract of feedstock production, and by the total alkali of this extract preparation.Described extract, total alkali and their preparation method can be referring to the patent of invention ZL00106234.4 of Nankai University and Inner Mongol University of Technology's co-applications, and the full text that this paper introduces this patent as a reference.With above-mentioned Cynanchun komarovii extract, or be referred to as " former medicine " by the total alkali of this extract preparation in description of the invention, they had both comprised extract, also comprised the total alkali of extract through making after separating.
For example, described Cynanchun komarovii extract can prepare as follows:
1, gets the Cynanchun komarovii acrial part natural air drying of harvesting early winter, be ground into fibrous;
2, processing industry ethanol covers forage degree of being with solution, in stirring at normal temperature lixiviate Separation of Solid and Liquid after 2-4 hour, repeats twice of above-mentioned steps; The determining alcohol of wherein said industrial alcohol is 60-95% normally;
3, merge extract, carry out decompression distillation, to reclaiming the about 1/2-2/3 of solvent, obtain the black slurries of pH＜5, i.e. the former medicine of Cynanchun komarovii extract.
And for example, the total alkali of described Cynanchun komarovii extract can prepare as follows:
1, the Cynanchun komarovii extract that said method is obtained transfers to PH＞8 with ammoniacal liquor, uses chloroform re-extract 3-6 time, and its Central Plains medicine is 7: 1 with the volume parts ratio of chloroform;
2, merge each batch extract, distillating recovering solvent is about 2/3, obtains the brown slurries, promptly former medicine total alkali; Also can be with preceding extract concentration extraction total alkali several times, will after several times extract directly recycle as extractant.
Above-mentioned Cynanchun komarovii extract or total alkali can directly be used as the raw material of antivirotic of the present invention without purifying, for improving effective component content can be used as antivirotic after purifying again raw material.
Preferably, the former medicine of Cynanchun komarovii extract described in the antivirotic of the present invention is the Cynanchun komarovii extract, and the content of 7-demethoxytylophorine is about 0.03-0.4% (weight) in the described extract.
Preferably, the former medicine of Cynanchun komarovii extract described in the antivirotic of the present invention is a Cynanchun komarovii extract total alkali, and the content of 7-demethoxytylophorine is about 0.1-5% (weight) in the described total alkali.
The auxiliary agent that is contained in the plant source disease-resistant toxic agent of the present invention is the assistant carrier of pesticidal preparations field routine, can select according to prepared formulation, described auxiliary agent for example is excipient, filler, dispersant, suspending agent, disintegrant, preservative or surfactant etc.
Plant source disease-resistant toxic agent of the present invention can be formulated to and be any formulations of pesticide commonly used, for example aqua, emulsifiable concentrate, suspending agent, granule or wetting powder etc.To water baseization, ultramicronising, no dust and the development of sustained release aspect, wherein water baseization is one of main direction of agricultural chemicals development, so preferred aqua, emulsifiable concentrate or wetting powder at the modern formulations of pesticide.
When plant source disease-resistant toxic agent of the present invention was aqua, described aqua by volume percentage meter comprised following component: former medicine 0.1-75%, solvent 11-80% and auxiliary agent 12-22% are preferably former medicine 10-75%, solvent 11-80% and auxiliary agent 12-22%.
When plant source disease-resistant toxic agent of the present invention is emulsifiable concentrate, described emulsifiable concentrate comprises following component in percent by volume: former medicine is 0.1-50%, solvent 20-70%, with auxiliary agent 7-30%, being preferably former medicine is 10-50%, solvent 20-70% and auxiliary agent 7-30%.
Wherein said solvent is any one solvent that is selected from water, ethanol, methyl alcohol, acetonitrile, ethylene glycol, chloroform or ethyl acetate, or the combination of two or more solvents arbitrarily.
Wherein said auxiliary agent is to be selected from one or more following assistant agents: surfactant, wetting agent, dispersant, suspending agent, filler, cosolvent, aromatic, colouring agent and preservative.
When preparation aqua or emulsifiable concentrate, the use of surfactant is extremely important.Concrete, surfactant used in the present invention comprises anion surfactant or non-ionic surface active agent, they can use separately, or one or more similar or inhomogeneous surfactant compounds use.For example various non-ionic surface active agents use separately or the composite use of several non-ionic surface active agents; Or various anion surfactants use separately or the composite use of several non-ionic surface active agents; Or one or more anion surfactants and the composite use of one or more non-ionic surface active agents.Wherein said non-ionic surface active agent for example is: the stearic acid isopropyl ester, emulsifier NR-40, the secondary alcohol APEO, rosin amine polyoxyethylene (2) ether, glutamic acid bay alcohol ester, double long-chain alkyl phenol polyethenoxy ether citrate, castor oil polyoxyethylene (20) ether, CGN-3 botanical pesticide surfactant, surfactant JS-PS007, amino-polyether organic silicon surfactant SSI (epoxy modified polysiloxane) and agricultural silicon surface active agent BD-3077.Described anion surfactant for example is C12～C16 secondary alcohol sodium sulfonate, oleic acid single ethanol amide-polyoxyethylene acid sodium sulphate, 2-tetradecyloxyaniline sodium propionate and succinic acid two (2-methyl pentyl ester) sodium sulfonate.
Also can use existing compound surfactant on the market, for example by non-ionic surface active agent and the composite commodity that form of anion surfactant: OX series pesticide micro emulsion aqueous emulsion emulsifier, trade name is compound OX-0657, English name microemulsifier for pesticideoxseries; And penetrating agent JFC (M type) (penatrant JFC (M type), they can use separately or with the composite use of other agricultural chemicals assistant agent.The compounded surfactant of selling on other market comprises the wetting agent JS-RDA dispersant (dispersant JS-RDA) and the dispersant HY-DC (dispersant HY-DC) of wetting powder.
Preferably, described surfactant is selected from any one of alkylphenol polyoxyethylene OP-10 or polyoxyethylene nonylphenol ether (n=12), castor oil and ethylene oxide condensate EL-40, fatty alcohol-polyoxyethylene ether AEO (10-15) or secondary alkanol APEO JFC, or two or more combination arbitrarily.
When plant source disease-resistant toxic agent of the present invention is wetting powder, described wetting powder comprises following component by weight percentage: former medicine 0.1-45%, wetting agent 8-22% and filling additive 33-91% are preferably former medicine 10-45%, wetting agent 8-22% and filling additive 33-80%.
Wherein said wetting agent can be the conventional wetting agent that uses, for example dodecyl sodium sulfate and/or draw back powder; Described filler can be the conventional filler that uses, for example the combination of any one of diatomite, talcum powder, atlapulgite and sodium lignin sulfonate or two kinds.
Usually, use or the transportation storage for convenient, can at first be prepared into the higher preparation of concentration, for example activity component concentration is greater than 0.03% (weight ratio) or the higher preparation of concentration, for example make the preparation that activity component concentration is 0.3%-5%, before using, obtain the preparation that activity component concentration is suitable for using, for example preparation of 0.0007%-0.01% (being 7-100ppm) by dilution.
Another purpose of the present invention has provided the preparation method of above-mentioned antivirotic, and this method can adopt the common method in pesticidal preparations field to carry out.
Concrete, for example when the aqua of preparation plant source disease-resistant toxic agent of the present invention, former medicine, solvent can be mixed by following volume ratio with auxiliary agent: former medicine: solvent: auxiliary agent=0.1-75: 11-80: 12-22, preferred former medicine: solvent: auxiliary agent=10-75: 11-80: 12-22, and preparation is promptly finished at room temperature abundant stirring.
When the emulsifiable concentrate of preparation plant source disease-resistant toxic agent of the present invention, with former medicine, solvent and auxiliary agent by following volume ratio than mixing: former medicine: solvent: auxiliary agent=0.1-50: 20-70: 7-30, preferred former medicine: solvent: auxiliary agent=10-50: 20-70: 7-30 and at room temperature fully stirring promptly finish preparation.
When the wetting powder of preparation plant source disease-resistant toxin preparation of the present invention, can at first dry former medicine, through mechanical crushing, former medicine after will pulverizing then evenly mixes by following weight ratio with wetting agent and filler: former medicine: wetting agent: filling additive=0.1-45: 8-22: 33-91, preferred former medicine: wetting agent: filling additive=10-45: 8-22: 33-80, and then crushing screening is promptly finished preparation.
Another purpose of the present invention has provided the application of above-mentioned plant source disease-resistant toxic agent, and described application comprises caused plant disease when being used to prevent and treat the virus host plant.
Concrete, the application of above-mentioned plant source disease-resistant toxic agent is that described antivirotic is used to prevent and treat one or more plant diseases that single virus infection different host or the integrated control same host of multiple different virus mixed infection or similar host cause.
In the present invention, described virus mainly is meant five kinds of viruses of Tobamovirus (TMV), Cucumovirus (CMV) and three Tobamovirus of Potyvirus (PVY): cucumber mosaic virus (CMV), tobacco mosaic virus (TMV), marmor upsilon (PVY), overgrown with weeds blue and white mosaic virus (TuMV) or Tomato mosaic virus (ToMV).
The host that virus mentioned above is contaminated mainly is meant plant, plant described in the present invention is the notion of general reference ordinary meaning, the present invention is applicable to all plants, be specially adapted to tobacco, vegetables, fruit crop and such as ornamental plants of flowers class etc. etc., especially tobacco such as spinach, crowndaisy chrysanthemum, shepherd's purse, leaf mustard etc.
Plant disease mentioned above is meant the disease that the virus dip-dye is caused, comprise the plant disease that same host of single virus infection or similar host cause, for example infect the plant disease that same host or similar host cause by cucumber mosaic virus (CMV), tobacco mosaic virus (TMV), marmor upsilon (PVY), overgrown with weeds blue and white mosaic virus (TuMV) or Tomato mosaic virus (ToMV).Described disease also comprises by two kinds in the above-mentioned virus or the caused plant disease of the same or similar host plant of multiple virus infection.
Virus is extensively to be present in natural world, and the host is in the obligate molecule parasite of life living cells.Mainly surround the virion that nucleic acid is formed no cell structure by protein.Every kind of virus has only a kind of nucleic acid type, and its nucleic acid replication and protein synthesis need rely on the host raw material and place are provided, the no dubbing system of virus self.The title plant virus of parasitic plant, zooparasitic title animal virus, the title phage of parasitic bacteria.The plant virus of having studied at present and having named reaches kind more than 1000, endangers serious plant virus again and belongs to and have six kinds in that world wide is widely distributed: 1, Cucumovirus (Cucumovirus); 2, Tobamovirus (Tobamovirus); 3, Potyvirus (Potyvirus); 4, yellow syndrome virus belongs to; 5, Nepovirus; With 6, the fungi baculoviral belongs to.The application of plant source disease-resistant toxic agent of the present invention relates to first three three type sepecies of planting Tobamovirus, and the summary situation is as follows:
1, CMV is the type sepecies of Cucumovirus (Cucumovirus)
It is in all viruses parasitic plant the most widely, CMV can infect and comprise unifacial leaf, dicotyledonous 67 sections, 470 various plants, and normal and other viral mixed infection.As: CMV and melon viruses (MMV) but 39 section's 117 plant species of mixed infection.CMV can survive the winter in the ornamental plants bodies such as flowers at perennial herb.Can rely on aphid to pass poison, juice passes poison, and host's seed passes poison.
2, TMV is the type sepecies of Tobamovirus (Tobamovirus)
Be distributed in all over the world, the host is extremely wide, and is strong to the environment resistance, can survive in dry blade more than 50 year.Main by the juice transmitted virus, as: in the farming operation transplanting is arranged, training, prune pinching, kidnapping, grafting, layer is planted and is used the vegetative propagation stem tuber, bulb, bulb etc., virus is promptly from the microtrauma mouth of plant corpus, in juice enters host plant body, can cause tobacco, tomato, capsicum, the mosaic disease of crops such as fruit tree.Grow with the host, virus relies on the host and carries out self-replacation and a large amount of propagation toxin rapidly, causes serious quality of host plant and production loss, even dead.
3, PVY is the type sepecies of Potyvirus (Potyvirus)
Be maximum in a plant virus genus, comprise 75 kinds.The PVY host range is wider, can infect Solanaceae, Chenopodiaceae, and 60 various plants of pulse family pass poison by aphid, and juice passes poison, and seed and asexual propagation material pass poison.
In order to prove the control efficiency of the plant disease that antivirotic of the present invention causes virus, adopt the method for indoor biometrics sample, tested the control efficiency of antivirotic of the present invention, optimized the aqua, emulsifiable concentrate and the wetting powder that filter out this plant source disease-resistant toxic agent five kinds of viral CMV, TMV of above-mentioned three Tobamovirus, PVY, TuMV, ToMV; Also find the inhibition effect difference of different vaccination ways to virus: as, medicament is mixed the back inoculation with viral juice, the drug effect that suppresses virus is 70-90%; Earlier the spray medicine inoculated after 24 hours, was 40-50% to the inhibition drug effect of virus; The difference of the two is that the former is that this anti-virus formulation directly acts on five kinds of viruses, enters the different host plant afterwards again, and having the poison that extremely significantly directly goes out is 70-90% with the dual preventive effect that suppresses virus replication in the different host plant corpus; The latter is sprayed medicine earlier, inoculates after 24 hours, and quite the preventive effect that host plant absorbs this anti-virus formulation earlier and transmission only has inhibition virus replication in the host plant body in plant corpus is 40-50%.The serious living test in field that CMV, TMV, PVY virus mixed infection tobacco are taken place over the years is tested and in Shandong, use this antiviral aqua diluted concentration 30ppm, to transplant the land for growing field crops also the tobacco (NC89) behind the seedling in left and right sides 28-40 at the beginning of 5 months days, spray medicine 4 times, absorb this anti-virus formulation and transmission in vivo by host plant, suppress the mixing of three kinds of viruses at natural fully following antiviral aqua of condition and duplicate in the plant corpus of tobacco NC89 kind, its preventive effect also is 40-50%.Use antiviral similar aqua, respectively in Yunnan, the field trial in Guizhou, diluted concentration 50-100ppm, sprayed medicine three times the TMV disease phase tobacco (K326) of starting seedling stage in 21-30 days in June, reaches more than 41.5% suppressing the preventive effect that the single virus of TMV contaminates tobacco (K326).
Surround the virion that nucleic acid is formed no cell structure at virus by protein.Every kind of virus has only a kind of nucleic acid type, virus can only rely on transcribing of host's biology and the translation system is carried out self-replacation, so application of this antivirotic, be valued for the influence that is not subjected to host's category different with viral kind, except that effectively single virus of control or integrated control more than two kinds the viral mixed infection host plant, also possesses the application prospect of effective control virus host bacterium, host animal, host's human body cell.
Plant source disease-resistant toxin preparation of the present invention is mainly used in the virus disease on control CMV, TMV, PVY, TuMV or ToMV virus host tobacco, the vegetable melon and fruit plant.For example, comprise 68 kinds of vegetable virus diseases of following 12 big classes by vegetables and propagation such as the blue and white mosaic virus of grassland (TUMV), cucumber mosaic virus (CMV), tobacco mosaic virus (TMV) or radish mosaic virus (RMV) host spinach, crowndaisy chrysanthemum, shepherd's purse, leaf mustard.
The first, 7 kinds of virus diseases of broccoli class vegetables: 1, broccoli virus disease (Broccoli virusdisease); 2, purple broccoli virus disease (red cabbage virus disease); 3 savoy virus diseases (savoy cabbage virus disease), 4, kale virus disease (collard virus disease); 5, bulb broccoli virus disease (kohlrabi virus disease); 6, spore broccoli virus disease (Brusselssprout virus disease); 7, cabbage mustard virus disease (Chinese kale virus disease);
The second, 5 kinds of virus diseases of Chinese cabbage group: 1, Chinese cabbage virus disease (pak-choi virusdisease); 2, cabbage heart virus disease (mock pad-choi virus disease); 3, green moss virus disease (Taicaivirus disease); 4, leaf mustard virus disease (Leaf mustard virus disease); 5, Wuta-tsai virus disease (Wuta cai virus disease);
The 3rd, 19 kinds of virus diseases of green vegetable class: 1, head lettuce virus disease (Head gardenlettuce virus disease); 2, cos lettuce virus disease (Longleaf garden lettuce virusdisease); 3, stem lettuce virus disease (Stem-eating lettuce virus disease); 4, witloof disease virus disease (Chicore virus disease); 5, crowndaisy chrysanthemum virus disease (Carland chrysanthemumvirus disease); 6, Gynura bicolor virus disease (Gynura virus disease); 7, watercress virus disease (Water cress virus disease); 8, capital water dish (Jingshuicai virus disease); 9, celery virus disease (Celery virus disease); 10, Japanese cryptotaenia stem and leaf virus disease (Mitsuba virus disease); 11, spinach virus disease (Spinach virus disease); 12, the leaf dish virus disease (Swiss chardvirus disease) of being unworthy of the honour; 13, the certain herbaceous plants with big flowers virus disease (Malabar spinach virus disease) that falls; 14, Bulbilus boussingaultiae pseudobaselloidis virus disease (Madeira-ving virus disease); 15, harmony dish virus disease (Water spinach virusdisease); 16, three-coloured amaranth virus disease (Edible amaranth virus disease); 17, Chinese mallow virus disease (Curled mallow virus disease); 18, New Zealand spinach virus disease (New Zealand spinachvirus disease); 19, long-life vegetable virus disease (Leaf-eating sweet potata virus disease);
The 4th, 8 kinds of virus diseases of melon vegetables and fruits: 1, little custard squash virus disease (Small custardsquash virus disease); 2, jade plate melon virus disease (Tray custard squash virus disease); 3, balsam pear virus disease (Bal sampear virus disease); 4, mini cucumber mosaic viral disease (Slicingcucumber mosaic virus disease); 5, trichosanthes anguina virus disease (Snake gourd virus disease); 6, watermelon virus disease (Water-melon virus disease); 7, melon viruses disease (Melon virusdisease); 8, pumpkin mosaic virus (squash mosaic virus disease);
The 5th, 6 kinds of virus diseases of solanaceous vegetables: 1, cherry and tomato ferm-leaf viral disease (Cherrytomato fern-like leaf virus disease); 2, cherry and tomato spotted wilt (Cherry tomato spotwilt virus disease); 3, cherry and tomato cecospora spot viral disease (Cherry tomato streak virusdisease); 4, cherry and tomato mosaic virus (Cherry tomato mosaic virus disease); 5, colored bell pepper virus disease (Ball pepper virus disease); 6, amorous eggplant (Penino virusdisease);
The 6th, 4 kinds of virus diseases of beans vegetables dish: 1, broad bean wilt virus disease (Broad bean wiltvirus disease); 2, garden pea, Lathyrus odoratus virus disease (Garden pea and sweet pea virusdisease); 3, French beans virus disease (lablab virus disease); 4, winged bean virus disease (Wingedbean virus disease);
The 7th, 3 kinds of virus diseases of root vegetables virus: 1, cherry radish, ternip virus disease (Cherry radish and Garden radish virus disease); 2, carrot virus disease (Carrotvirus disease); 3, root dish (filiale) virus disease (Table beet virus disease) of being unworthy of the honour;
The 8th, 3 kinds of virus diseases of tuber and tuberous rooted vegetables: 1, potato virus disease (Potato virusdisease); 2, the sick virus disease (Taro virus disease) of taro; 3, burnt taro virus disease (Edible cannavirus disease);
The 9th, 4 kinds of virus diseases of green onion garlic class and aromatic series vegetables: 1, peppermint virus disease (Mint virusdisease); 2, carolus disease viral disease (Sweet basil virus disease); 3, fennel virus disease (Fennelvirus disease); 4, coriander virus disease (Coriander virus disease);
The tenth, 3 kinds of virus diseases of other greengrocery virus: 1, edible mushroom virus disease (Ediblechrysanthemum virus disease); 2, Chinese artichoke virus disease (Chinese artichoke virusdisease); 3, gumbo virus disease (Okra virus disease);
The 11,4 kinds of virus diseases of permanent vegetable class: 1, Chinese toon virus disease (Chinese toonvirus disease); 2, lycium virus disease (Chinese wolfberry virus disease); 3, balloonflower root virus disease (Balloon flower virus disease); 4, lily disease viral disease (Lily virus disease); With
The 12,2 kinds of virus diseases of edible mushroom greengrocery: 1, flat mushroom virus disease (Pleurotusostreatus virus disease); 2, mushroom virus (Agaricus bisporus virus disease).
Plant described in the present invention is meant the notion of ordinary meaning, and the plant that the present invention is suitable for using is meant tobacco, vegetable melon and fruit crop and ornamental plants etc. especially.But at the different plant of virus host, use the optium concentration of this antivirotic, need through give birth to test test be defined as appropriate.
Of the present invention using than depending on the multiple factor such as vegetative period, weather of type, the order of severity, the floristics of disease, different region, crop with application concentration, usually dispenser uses than counting every mu of 0.3-2.0g/15-40L water with the quality of active component 7-demethoxytylophorine, suitable application concentration is that the content of active component 7-demethoxytylophorine in the preparation is 7-100ppm, preferred 20-50ppm, more preferably 25-40ppm.For example in seedling phase of tobacco, the transfer field is the tobacco seedling behind the seedling also, NC89 tobacco seedling for example, and it is used than being every mu of 0.3-0.6g/15L water, suitable application concentration is that the content of 7-demethoxytylophorine is 20-40ppm, is preferably about 30ppm; But, it is used than being every mu of 0.7-2.0g/20L water seedling stage in the tobacco, and application concentration is that the content of 7-demethoxytylophorine is 35-100ppm, is preferably about 40ppm.
Preferably, bright through the living testing authentication of indoor biological test and field: various preparations of the present invention can be used for preventing and treating the dip-dye of single virus, and described single virus comprises marmor upsilon (PVY), cucumber mosaic virus (CMV), overgrown with weeds blue and white mosaic virus (TuMV) or Tomato mosaic virus (ToMV); But and the multiple virus of integrated control, for example by two kinds or the multiple dip-dye that is selected from the virus of tobacco mosaic virus (TMV), marmor upsilon (PVY), cucumber mosaic virus (CMV), overgrown with weeds blue and white mosaic virus (TuMV) and Tomato mosaic virus (ToMV) to plant; And because the disease that above-mentioned dip-dye causes on one or more plants, its integrated control effect can reach 40-50%.Prove that various preparation of the present invention is the virus disease that causes of the single plant species virus of control plant or effective preparation of the two or more virus diseases that cause.When particularly using it for multiple virus to the dip-dye of tobacco, for example in Shandong Province's sunshine and two places, Linyi, to the serious mark formula kind that three main Tobamovirus take place over the years, comprise marmor upsilon (PVY), tobacco mosaic virus (TMV) and cucumber mosaic virus (CMV) mix the field efficiency test of contaminating tobacco NC89 kind, use the antiviral aqua of basis of 30ppm concentration, to transplanting the also NC89 tobacco crop behind the seedling of land for growing field crops, in 28-40 days, spray medicine 4 times continuously, compare with two conventional medicines, product as strong your the biochemical technology development company in Heilungkiang, promptly 250 times of liquid of 2% bacterium gram poison gram dilute, i.e. effective ingredient Ningnanmycin concentration 80ppm; As Xian District, Shanxi Province wave chemical industry Co., Ltd product, promptly 3.95% virus must restrain 500 times of dilutions of wetting powder, and quite the effective ingredient virazole adds CuSO
4Add ZnSO
4Concentration be 79ppm.Later in the table 1,2,3 respectively behind this medicine of record spray the preventive effect after 50,66,75 days suitable with contrast medicine preventive effect.Low 2.6 times of above two the contrast medicines of the working concentration of this anti-virus formulation.Do not find that under this concentration this anti-virus formulation is to tobacco and other biological generation harmful effect.Result of the test shows that the control efficiency of antivirotic of the present invention obviously is better than the known antivirotic of prior art, has higher activity and safety.
Antivirotic acute oral toxicity LD of the present invention
50Female=hero＞5000mg/kg BW; Acute dermal toxicity LD
50Female=hero＞2000mg/kg.BW; All belong to low toxicity.Verify that thus anti-virus formulation of the present invention is a kind of harmless to people, animal, again botanical pesticide safely and efficiently.
Embodiment
Below by concrete embodiment technical scheme of the present invention is further described, the embodiment that wherein exemplifies only is to explanation of the present invention, and can not limit protection scope of the present invention by any way.
The preparation of the former medicine of embodiment 1 Cynanchun komarovii extract
1, gets the wild Cynanchun komarovii acrial part natural air drying of West Inner Mongolia of 150 kilograms of harvestings early winter, be ground into fibrous;
2, processing industry ethanol covers forage degree of being with solution, in stirring at normal temperature lixiviate Separation of Solid and Liquid after 2-4 hour, repeats twice of above-mentioned steps;
3, merge extract, carry out decompression distillation,, obtain the black slurries of 18 liters of pH＜5 to reclaiming the about 1/2-2/3 of solvent, i.e. the former medicine of Cynanchun komarovii extract, density is greater than 1, and wherein the content of 7-demethoxytylophorine is about 0.32%.
The preparation of embodiment 2 total alkalis
The Cynanchun komarovii extract that embodiment 1 obtains is transferred PH＞8 with ammoniacal liquor, in separatory funnel with chloroform re-extract 6 times, last chloroform extraction fluid point sample on lamellae, spray Dragerdorf reagent behind the chromatography, do not show salmon patch, the expression alkaloid in the most extract that come together; Former medicine is 7: 1 with the volume parts ratio of chloroform, merges first three batch extract, and it is about 2/3 to reclaim solvent, pours out the brown slurries, i.e. the former medicine of total alkali (1), and wherein the content of 7-demethoxytylophorine is 0.55%.The three batches of extracts in back are directly applied mechanically again as extractant, and the content that reclaims the former medicine of total alkali (2) behind the solvent at last is 0.27%.
The preparation of embodiment 3 aquas of the present invention
In modulation kettle, add embodiment 1 preparation former medicine 12.0L, 2.0L acetonitrile and 2.5L OP-10 (Beijing radically reform protect bio tech ltd provide) are provided, stir 40min at normal temperatures, make this aqueous pesticide solution of 16.5L, the content of 7-demethoxytylophorine is 0.26% in this aqua.
The preparation of embodiment 4 emulsifiable concentrates of the present invention
Get total alkali (1) 110ml of embodiment 2 preparations, 100 milliliters of acetonitriles and 16ml OP-10 (Beijing radically reform protect bio tech ltd provide) are provided, stir 40min at normal temperatures, promptly make the 226ml emulsifiable concentrate, the content of 7-demethoxytylophorine is 0.34% in this emulsifiable concentrate.
The preparation of embodiment 5 wetting powders of the present invention
Get the total alkali (2) that embodiment 2 makes, put into oven for drying, pulverize, get dry former medicine 28 grams, wherein the content of 7-demethoxytylophorine is 0.43%.Itself and wetting agent dodecyl sodium sulfate 6g and filler atlapulgite 41g are stirred 20min at small size spice machine central roll, carry out mechano-electronic mill processing 20min, cross 300 mesh sieves, promptly make this pesticide wettable of 75g, the content of 7-demethoxytylophorine is 0.16% in this wetting powder.
Embodiment 6 indoor living body biological tests
When the content that present embodiment has been tested 7-demethoxytylophorine in the antivirotic of the present invention is 20ppm to the control efficiency of 5 kinds of viruses.
1. test objective: when this antivirotic sample is 20ppm at the content of 7-demethoxytylophorine, measure preventive effect, optimize the prescription that filters out aqua, emulsifiable concentrate, wetting powder to 5 kinds of viruses by the withered spot method of indoor live body.Prove that simultaneously this anti-virus formulation sample infects the different host plant to five kinds of virus is single, realize viral self-replacation to relying on the host, this anti-virus formulation all has significantly extensively inhibitory action.
2. test material:
Virus/test plant: TMV/ Nicotiana glutinosa, the yellow leaf cigarette of PVY/, CMV/ Nicotiana glutinosa, TuMV/ Chenopodium amaranticolor, ToMV/ Nicotiana glutinosa;
For aqua, emulsifiable concentrate and the wetting powder of examination antivirotic: embodiment 3,4 and 5, the content that is diluted to the 7-demethoxytylophorine is 20ppm.
3. test method:
After the sick vanes that infects the test plant of TMV, PVY, CMV, TuMV, ToMV ground, with this antivirotic sample mixed, with the dilution of inoculation buffer solution, make each sample solution concentration all reach 20ppm, be inoculated in respectively behind the mixing in the strain of test plant, every strain connects the 3-5 leaf, every leaf 2-3 square centimeter, if the clear water contrast, withered spot number on the statistics blade after 3-5 days.Calculate control efficiency according to following formula.
The antiviral aqua of the present invention, emulsifiable concentrate, wetting powder are target with five kinds of viruses (TMV, CMV, PVY, TuMV and ToMV), and the living survey of indoor live body control efficiency was as follows when drug concentration of the present invention was 20ppm:
The field control effect test of embodiment 6 antivirotics of the present invention
2004-2005, in Rizhao City Mao Bu town, Shandong Province and Yang Po town, Linyi City Yinan County, the aqua of using plant source disease-resistant toxic agent of the present invention has carried out the control efficiency test in field.
1. experimental condition
1.1 crop and target crop are tobacco bred NC89: target is tobacco mosaic virus (TMV), cucumber mosaic virus (CMV), marmor upsilon (PVY)
1.2 experimental scheme and processing are slow than chemical pesticide because of botanical pesticide performance drug effect, ad hoc meter experimental program of the present invention.Adopt district's group arrangement at random, repeat 3 times, every sub-district plants cigarette 100 strains, line-spacing 1.6m, sub-district area 72m
2Use manual sprayer also to begin dispenser behind the seedling from transplanting the land for growing field crops, 7-10 days 1 time, continuous 4 times, to carry out in strict accordance with code requirement during dispenser, the concentration of making up a prescription is accurate, and leave dual sides is evenly thoughtful during the spray medicine.
2. result of the test
Result of the test sees Table 1-3.
Table 1 has write down and in 66 days, had extracted 50 strains by 5 samplings and carry out investigation statistics 3 times after medicament medicine of the present invention is used in Linyi in 2004, has reported 42 data; Table 2 has write down and in 50 days, had extracted 50 strains by 5 samplings and carry out investigation statistics 3 times after the Shandong Province uses medicament of the present invention sunshine in 2004, reports 23 data; Table 3 has write down the experimental condition that repeated sunshine in 2005, behind medicine in 75 days, extracts the preventive effect data that investigation statistics 1 time is carried out in 50 strains in onset peak period (about mid-July) by 5 samplings." the tobacco diseases grade scale " formulated according to China Tobacco Corporation carries out classification, calculates the incidence of disease and disease index, calculates the control efficiency of various processing with disease index.
The grade scale of tobacco virus disease (YC/T 39-1996) is according to the classification of disease severity:
0 grade: complete stool is anosis:
1 grade: lobus cardiacus veinclearing or slight floral leaf, or top 1/3rd blade floral leaves are but indeformable, and plant does not have obvious dwarfing;
2 grades: 1/3rd to 1/2nd blade floral leaves, or minority deformable blade, or master pulse blackening, plant are downgraded and are more than 2/3rds of normal plant height;
3 grades: 1/2nd to 2/3rds blade floral leaves, or distortion, or the necrosis of master arteries and veins, or the plant dwarfing is 2/3rds to 1/2nd of normal plant height;
4 grades: complete stool blade floral leaf, gross distortion or necrosis, diseased plant downgrade for the normal plant height more than 1/2nd to 1/3rd.
The computational methods of the incidence of disease, disease index and control efficiency:
3 conclusions
3.1 from the field test results in 2 years as can be seen, contain the antiviral aqua of plant resource 7-demethoxytylophorine, the concentration of active ingredient is 30 * 10
-6G/mL (30ppm), integrated control effect to PVY, CMV and TMV when every mu amount of application is counted 0.3g with the quality of active component 7-demethoxytylophorine is 40%-50%, and (the active ingredient virazole adds CuSO must to restrain 500 times of liquid with conventional dose 3.95% virus of using in the present production
4Add ZnSO
4Concentration be 79 * 10
-6G/mL), (active ingredient Ningnanmycin concentration is 80 * 10 to 250 times of liquid of 2% bacterium gram poison gram
-6G/mL) control efficiency close (seeing Table 1,2,3), but the working concentration of the antiviral aqua of the present invention is than low 2.6 times of the working concentration of virazole or Ningnanmycin.
3.2 under the concentration of being used, antiviral aqua of the present invention growing and other biological generation harmful effect to tobacco do not found in the field.
The field test results of 2004 and 2005 sees table 1-3, and term " disease refers to " used in the table is the abbreviation of " disease index ", shows that antivirotic suppresses the replication capacity of single virus in the tobacco body; Used term in the table " comprehensive disease refers to ", promptly " comprehensive disease index " shows that antivirotic suppresses the replication capacity of three kinds of viruses in the tobacco body.
The field efficacy of table 3 antivirotic of the present invention is measured (2005, sunshine)
Repeat the field trial of aqua of the present invention sunshine in Shandong, formulation concentrations 30ppm, condition is with table 2, and investigation in 75 days once reports that multiple viral TMV, CMV and PVY mix the preventive effect report of contaminating tobacco after medication
Annotate: aqua concentration of the present invention is 30ppm; Medicament virus must restrain and be meant that 3.95% virus must restrain 500 times of wetting powder (Xian District, Shanxi Province wave chemical industry Co., Ltd) dilutions, quite 79ppm.
Preceding twice date of record is the data in 50 days behind the medicine in the table 1, and wherein the disease of aqua of the present invention refers to or comprehensive disease refers to all to show that smaller or equal to the contrast medicine ability that suppresses virus is greater than the contrast medicine; The data verification of table 2 above-mentioned feature.In the table 1 disease on July 15 refer to or comprehensive disease refer to be worth be the value of aqua of the present invention greater than the contrast medicine, disclosed the ability that suppressed virus behind the aqua spray medicine of the present invention in 66 days and transferred the feature that is lower than the contrast medicine to.Table 3 is single-round survey data that same test conditions is put in poison back 75 days, and comprehensive disease refers to be lower than the contrast medicine and verified that aqua spray medicine of the present invention suppressed the fact that viral ability is lower than the contrast medicine slightly in 66 days.
According to the listed field test results of above table 1-3, illustrate that antivirotic of the present invention is effective preparation of control a kind of viral TMV of tobacco and multiple viral TMV CMV of comprehensively preventing and controlling tobacco and PVY dip-dye.Drug effect 40-50% is close for its control efficiency and conventional dose (virus must restrain, the bacterium gram is malicious restrains), and not seeing has harmful effect to other plant.But the working concentration 30ppm of medicament of the present invention comparison is according to low 2.6 times of medicament (contrast drug concentration 80ppm), explanation is when same concentrations, the control efficiency of antivirotic of the present invention is higher than antivirotic commonly used far away, and because antivirotic of the present invention derives from plant itself, therefore having better safety, is natural, efficient, safe plant source disease-resistant poison agricultural chemicals system.
Below described embodiment of the present invention in detail, can do a lot of improvement and variation obviously for a person skilled in the art and can not deviate from essence spirit of the present invention.All these changes and improvements are all within protection scope of the present invention.
Claims (27)
1, a kind of plant source disease-resistant toxic agent, wherein containing with the 7-demethoxytylophorine is that the former medicine of Cynanchun komarovii extract of active ingredient is an active component, in the gross weight of antivirotic, the content of described former medicine is 0.1-75%, and the content of auxiliary agent is 99.9-25%; Wherein said former medicine is Cynanchun komarovii extract or Cynanchun komarovii extract total alkali, and wherein the content of 7-demethoxytylophorine is 0.03-5% (weight), and the pH of described Cynanchun komarovii extract＜5; Wherein said antivirotic is aqua, emulsifiable concentrate or wetting powder.
2, according to the described antivirotic of claim 1, when the former medicine of wherein said Cynanchun komarovii extract was the Cynanchun komarovii extract, wherein the content of 7-demethoxytylophorine was 0.03-0.4% (weight).
3, according to the described antivirotic of claim 1, when the former medicine of wherein said Cynanchun komarovii extract was Cynanchun komarovii extract total alkali, wherein the content of 7-demethoxytylophorine was 0.1-5% (weight).
4, according to any described antivirotic of claim 1-3, wherein said antivirotic is an aqua, and by volume the described aqua of percentage meter comprises following component: former medicine 0.1-75%, solvent 11-80% and auxiliary agent 12-22%.
5, antivirotic according to claim 4, wherein said aqua by volume percentage meter comprise following component: former medicine 10-75%, solvent 11-80% and auxiliary agent 12-22%, condition is that each component sum is 100%.
6, according to any described antivirotic of claim 1-3, wherein said antivirotic is an emulsifiable concentrate, comprises following component in the described emulsifiable concentrate of percent by volume: former medicine is 0.1-50%, solvent 20-70% and auxiliary agent 7-30%.
7, antivirotic according to claim 6, wherein said emulsifiable concentrate comprises following component in percent by volume: former medicine is 10-50%, solvent 20-70% and auxiliary agent 7-30%.
8, according to claim 5 or 7 described antivirotics, wherein said solvent is selected from any one of water, ethanol, methyl alcohol, acetonitrile, ethylene glycol, chloroform and ethyl acetate, or wherein two or more combination arbitrarily.
9, according to claim 5 or 7 described antivirotics, wherein said auxiliary agent is selected from one or more following adjuvants: surfactant, wetting agent, dispersant, suspending agent, filler, cosolvent, aromatic, colouring agent and preservative.
10, antivirotic according to claim 9, wherein said surfactant is selected from any one among alkylphenol polyoxyethylene OP-10 or polyoxyethylene nonylphenol ether (n=12), castor oil and ethylene oxide condensate EL-40, fatty alcohol-polyoxyethylene ether AEO (10-15) and the secondary alkanol APEO JFC, or wherein two or more combination arbitrarily.
11, according to any described antivirotic of claim 1-3, wherein said antivirotic is a wetting powder, and described by weight percentage wetting powder comprises following component: former medicine 0.1-45%, wetting agent 8-22% and filling additive 33-91%.
12, antivirotic according to claim 11, wherein said wetting powder comprises following component by weight percentage: former medicine 10-45%, wetting agent 8-22% and filling additive 33-80%.
13, antivirotic according to claim 12, wherein said wetting agent are dodecyl sodium sulfate and/or draw back powder; Described filling additive is any one or any two kinds combination that is selected from diatomite, talcum powder, atlapulgite and sodium lignin sulfonate.
14, according to the preparation method of any described antivirotic of claim 1-13, described antivirotic can adopt the conventional method preparation in pesticidal preparations field.
15, according to the application of any described antivirotic of claim 1-13, described application comprises caused plant disease when being used to prevent and treat the virus host plant, and wherein said virus is following three kinds of Tobamovirus: five kinds of viruses of Tobamovirus (TMV), Cucumovirus (CMV) and Potyvirus (PVY): cucumber mosaic virus (CMV), tobacco mosaic virus (TMV), marmor upsilon (PVY), overgrown with weeds blue and white mosaic virus (TuMV) or Tomato mosaic virus (ToMV).
16, according to the application of the described antivirotic of claim 15, wherein said virus is the virus of Cucumovirus (CMV).
17, according to the application of the described antivirotic of claim 15, described antivirotic is used to prevent and treat one or more plant diseases that single virus infection different host or the integrated control same host of multiple different virus mixed infection or similar host cause.
18, the application of antivirotic according to claim 17, wherein said virus is contaminated on tobacco or vegetable melon and fruit plant.
19, the application of antivirotic according to claim 18, wherein said virus is contaminated on tobacco.
20, according to the application of any described antivirotic of claim 15-19, using than the weight in active ingredient 7-demethoxytylophorine of wherein said antivirotic is every mu of 0.3-2.0g/15-40L water.
21, according to the application of the described antivirotic of claim 20, the using of wherein said antivirotic than weight in active ingredient 7-demethoxytylophorine, to the transfer field seedling of the tobacco behind the seedling also, it is used than being every mu of 0.3-0.6g/15L water.
22, according to the application of the described antivirotic of claim 20, the using of wherein said antivirotic than weight in active ingredient 7-demethoxytylophorine, in the middle seedling stage of tobacco, it is used than being every mu of 0.7-2.0g/20L water.
23, according to the application of any described antivirotic of claim 15-19, the application concentration of wherein said antivirotic is 7-100ppm.
24, according to the application of the described antivirotic of claim 23, wherein in the seedling phase of tobacco, its application concentration is 20-40ppm.
25, according to the application of the described antivirotic of claim 24, wherein in the seedling phase of tobacco, its application concentration is preferably 30ppm.
26, according to the application of the described antivirotic of claim 23, wherein in the middle seedling stage of tobacco, its application concentration is 35-100ppm.
27, according to the application of the described antivirotic of claim 26, wherein in the middle seedling stage of tobacco, its application concentration is preferably 40ppm.
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| CN102696413B (en) * | 2011-11-21 | 2014-06-04 | 云南省烟草公司昆明市公司 | Method for preventing and controlling tobacco mosaic diseases in field planting period of tobacco floating seedling raising |
| CN103960229A (en) * | 2013-01-25 | 2014-08-06 | 江苏蓝丰生物化工股份有限公司 | Tylophorine malate (NK-007) water aqua and preparation method thereof, and application of tylophorine malate water aqua in prevention and treatment of viral diseases of tobacco, capsicum and tomatoes |
| CN105130985B (en) * | 2015-09-25 | 2017-08-25 | 南开大学 | Phenanthroindolizididerivative pyridine alkaloid quaternary ammonium salt derivatives and its preparation and Antiphytoviral application |
| CN105706881A (en) * | 2016-03-16 | 2016-06-29 | 沈素香 | Little pepper fast propagation method |
| CN112450230A (en) * | 2020-11-03 | 2021-03-09 | 南京植创生物技术研究院有限公司 | Compound plant disinfectant and preparation method thereof |
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