CN101265241B - Method for preventing and curing plant disease by increasing plant immunity and use thereof - Google Patents

Abstract

The invention provides a plant protective method and application of inducing immune system of plant by 4-methyl-1,2,3-thiadiazolyl-5-formic acid (NK-F001), sodium 4-methyl-1,2,3-thiadiazolyl-5-formate (NK-F002), 4-methyl-1,2,3-thiadiazolyl-5-ethyl formate (NK-F003) and methenaimde (SZG-7) to generate resistance against the plant virus. The invention also provides an application for improving the plant immunity by combination of two or three of 4-methyl-1,2,3-thiadiazolyl-5-formic acid (NK-F001), sodium 4-methyl-1,2,3-thiadiazolyl-5-formate (NK-F002), 4-methyl-1,2,3-thiadiazolyl-5-ethyl formate (NK-F003), methenaimde (SZG-7), benzthiadiazole (activated ester, BTH), tiadinil (TDL), DL-Beta-aminobutyric acid, virazole, ningnanmycin, antofine, Bingduxing No.1, Bingduxing No.2, XY-13 and XY-30 so as to induce the plant to generate antiviral activity, processing dosage forms of single and mixture preparations, and compositions and contents of effective components in the dosage forms. The invention discloses a plant inducing range and plant antiviral inducing range of compounds and compositions thereof, and application of plant protection in the fields of agriculture and horticulture thereof.

Description

Utilize the raising via plant immunity to prevent and treat method of phytopathy murder by poisoning and uses thereof

Technical field

Technical scheme of the present invention relates to the method that the compound that utilizes chemosynthesis improves via plant immunity, and these compounds are improving the purposes that agricultural plants and gardening plant opposing virus disease infects.

Background technology

The viroses of plant are plant " cancers ", its method of preventing and treating mainly comprises the method for physics and the method for method one machinery of chemistry at present, but each method all can not reach the ideal effect, in chemical process, also there is not at present a kind of medicament that fully can the controlling plant virus disease, owing to there is relation closely between the generation of virus disease and the host plant, therefore, the control that exploration utilizes the mechanism of plant self resistance to carry out virus disease is agricultural sciences worker's the ideal and the direction of effort always, the plant activator is the outstanding achievement of this direction research just, it can avoid traditional agricultural chemicals to obtain insect protected by directly acting on insect and weeds and pathogen simultaneously, the purpose of control grass and diseases prevention and cause beneficial organism is caused detrimentally affect and to the pressure of environment, at worldwide widely-used (the Bakulev V A of plant activator BTH that exploitation is the most successful at present, Dehaen W.TheChemistry of 1,2,3-thiadiazole, 2004, John Wiley﹠amp; Sons, Inc.), the contriver is from fundamental research, in state natural sciences fund (30270883 and 20672062), under the subsidy of " 973 " plans (2003CB114402) and Tianjin natural science fund (07JCYBJC01200) and Tianjin science and technology support plan International Technology collaborative project (07ZCGHHZ01400), synthetic and discovery N-(the 5-methyl isophthalic acid of design, the 3-thiazol-2-yl)-the 4-methyl isophthalic acid, 2,3-thiadiazoles-5-methane amide (hereinafter to be referred as N-(5-methyl-1,3-thiazole-2-yl)-4-methyl-1,2,3-thiadiazole) has good biological activity (Fan Zhijin etc., novel [1,2,3] thiadiazoles derivative and preparation method and use thereof, People's Republic of China's national inventing patent, publication number: CN1810808A; Fan Zhijin etc. have the new compound of activity of inducing secondary metabolism of Hypericum plant, People's Republic of China's national inventing patent, publication number: CN2020667A); Thereafter, the contriver to this compounds carried out the composition optimizes of system and initiative research (Fan Zhijin, etc., thiobizole formanide derivative and preparation method and use thereof, People's Republic of China's national inventing patent, application number: 200810052546.6; Fan Zhijin, etc., the heterogeneous ring compound and the preparation method and use thereof of thiadiazoles, People's Republic of China's national inventing patent, application number: 200810052547.0 contained; Fan Zhijin, etc., thiadiazoles imine derivative and preparation method and use thereof, People's Republic of China's national inventing patent, application number: 200810052553.6).N-(5-methyl-1,3-thiazole-2-yl)-4-methyl-1,2,3-thiadiazole is to the LD of the acute oral of female, mouse great and mighty or powerful ₅₀All greater than 5000mg/kg; To acute dermal LD female, mouse great and mighty or powerful ₅₀All greater than 2000mg/kg; Acute eye irritation test result to rabbit is slight the stimulation; Acute skin irritation test result to rabbit is non-stimulated; Sensitization of skin test-results to cavy is weak sensitization.Originally study carefully invention and relate to method and the purposes of these compounds in agricultural plants and gardening plant protection that compounds such as utilizing N-(5-methyl-1,3-thiazole-2-yl)-4-methyl-1,2,3-thiadiazole improves via plant immunity.

Summary of the invention

Technical problem to be solved by this invention is: new 4-methyl isophthalic acid is provided, 2, the synthetic method of 3-thiadiazoles-5-sodium formiate, 4-is provided methyl isophthalic acid, 2,3-thiadiazoles-5-formic acid and 4-methyl isophthalic acid, 2,3-thiadiazoles-5-ethyl formate and N-(5-methyl-1,3-thiazole-2-yl)-4-methyl-1,2,3-thiadiazole improve the method for via plant immunity, and utilize these compounds to improve via plant immunity is used for the viral diseases of plants control in agriculture field and gardening field application.

The present invention solves this technical problem the technical scheme that is adopted: have the compound that improves via plant immunity and comprise the 4-methyl isophthalic acid, 2,3-thiadiazoles-5-formic acid and 4-methyl isophthalic acid, 2,3-thiadiazoles-5-ethyl formate and N-(5-methyl-1,3-thiazole-2-yl)-4-methyl-1,2,3-thiadiazole and 4-methyl isophthalic acid, 2,3-thiadiazoles-5-sodium formiate, its chemical structure sees Table 1:

The chemical structure of table 1 compound and test code number

Chemical structure

Test code number NK-F001 NK-F002 NK-F003 N-(5-methyl-1,3-thiazole-2-yl)-4-methyl-1,2,3-thiadiazole (SZG-7)

4-methyl isophthalic acid wherein, 2,3-thiadiazoles-5-formic acid (NK-F001), the 4-methyl isophthalic acid, 2,3-thiadiazoles-5-sodium formiate (NK-F002), the 4-methyl isophthalic acid, 2,3-thiadiazoles-5-ethyl formate (NK-F003) and N-(5-methyl-1,3-thiazole-2-yl)-4-methyl-1,2,3-thiadiazole (SZG-7) they can be the compounds of synthetic compound or purchase.Diazosulfide (Acibenzolar, BTH) and tiadinil (TDL) and DL-beta-aminobutyric acid, virazole, Ningnanmycin, antofine, No. 1, viral star, No. 2, viral star and compounds such as XY-13, XY-30 can synthesize or buy.

The method that compound among the present invention improves via plant immunity is as follows:

The mensuration of stripped direct antiviral activity adopts half leaf method to carry out; It is common cigarette with the seedling age unanimity that live body is induced, 3 basins are one group, cigarette seedling respectively at inoculation pre-treatment in preceding 7 days, processing mode comprises: spray test compound solution 2 to 3 times, each 10 milliliters, or soil treatment, each 10 milliliters, the 7th day frictional inoculation TMV on the tobacco leaf that newly grows, the cigarette seedling is placed its growth optimal temperature and illumination, and cultivation is after 3 days down, and inspection incidence, comprehensive scab number are calculated as follows out the inducing anti-disease toxic effect fruit of test compound to TMV, 3 repetitions are established in each processing, and contrast divides blank and standard chemicals treatment to contrast 2 kinds; The relative effect of test compounds is divided into 4 grades: A, B, C, D, and concrete data are the A level: induce effect＞70%, for excellent; B level: induce effect 70～50%, for very; C level: induce effect 30～50%, for poor; D level: induce effect＜30% to be considered as not having and induce effect:

$R=\frac{\mathrm{CK}-I}{\mathrm{CK}}\×100$

Wherein, R is the induce effect of new compound to the anti-TMV of tobacco, unit: %

CK is the average withered spot number of clear water contrast blade, unit: individual

I is for inducing the average withered spot number of handling rear blade, unit: individual through compound;

Same method can be used for these compounds and mix the use anti-TMV of evoking tobacco or other food crop and cash crop and gardening plant anti-plant viral disease and application thereof separately or with any 2 kinds or any three kinds.

The invention has the beneficial effects as follows: the present invention utilizes the 4-methyl isophthalic acid, 2,3-thiadiazoles-5-formic acid (NK-F001), the 4-methyl isophthalic acid, 2,3-thiadiazoles-5-sodium formiate (NK-F002), the 4-methyl isophthalic acid, 2,3-thiadiazoles-5-ethyl formate (NK-F003) and N-(5-methyl-1,3-thiazole-2-yl)-4-methyl-1,2,3-thiadiazole (SZG-7), diazosulfide (Acibenzolar, BTH) and tiadinil (TDL) and DL-beta-aminobutyric acid, virazole, Ningnanmycin, antofine, No. 1, virus star, No. 2, virus star and XY-13, compounds such as XY-30 or its composition can improve the immunizing power of agricultural plants and gardening plant, strengthen the invasion of agricultural plants and gardening plant plant opposing plant virus, be used for the control of the viroses of plant.

The present invention will more specifically describe by specific preparation and biological activity determination embodiment and utilize the 4-methyl isophthalic acid, 2,3-thiadiazoles-5-formic acid (NK-F001), the 4-methyl isophthalic acid, 2,3-thiadiazoles-5-sodium formiate (NK-F002), the 4-methyl isophthalic acid, 2,3-thiadiazoles-5-ethyl formate (NK-F003) and N-(5-methyl-1,3-thiazole-2-yl)-4-methyl-1,2,3-thiadiazole (SZG-7), diazosulfide (Acibenzolar, BTH) and tiadinil (TDL) and DL-beta-aminobutyric acid, virazole, Ningnanmycin, antofine, No. 1, virus star, No. 2, virus star and XY-13, compound such as XY-30 and composition thereof improve method and the biological activity and the application thereof of agricultural plants and gardening plant Antiphytoviral disease, but described embodiment only is used to specify the present invention and unrestricted the present invention, and embodiment is as follows:

Embodiment 1

Utilize NK-F001, NK-F002, NK-F003 and SZG-7 etc. improve method and the purposes of the immunizing power opposing TMV of tobacco

The mensuration of stripped direct antiviral activity adopts half leaf method, it is common cigarette with the seedling age unanimity that live body is induced, 3 basins are one group, cigarette seedling respectively at inoculation pre-treatment in preceding 7 days, processing mode comprises: spray test compound solution 2 to 3 times, each 10 milliliters, or soil treatment, each 10 milliliters, the 7th day frictional inoculation TMV on the tobacco leaf that newly grows places its growth optimal temperature and the following cultivation of illumination after 3 days the cigarette seedling, check incidence, comprehensive scab number is calculated as follows out the inducing anti-disease toxic effect fruit of test compound to TMV, and 3 repetitions are established in each processing, and contrast divides blank and standard chemicals treatment to contrast 2 kinds; The relative effect of test compounds is divided into 4 grades: A, B, C, D, and concrete data are the A level: induce effect＞70%, for excellent; B level: induce effect 70～50%, for very; C level: induce effect 30～50%, for poor; D level: induce effect＜30% to be considered as not having and induce effect:

$R=\frac{\mathrm{CK}-I}{\mathrm{CK}}\×100$

Wherein, R is the induce effect of new compound to the anti-TMV of tobacco, unit: %

CK is the average withered spot number of clear water contrast blade, unit: individual

I is for inducing the average withered spot number of handling rear blade, unit: individual through compound;

The mensuration of stripped direct antiviral activity adopts the half leaf method measurement result to show, the 4-methyl isophthalic acid, 2,3-thiadiazoles-5-formic acid (NK-F001), 4-methyl isophthalic acid, 2,3-thiadiazoles-5-sodium formiate (NK-F002), 4-methyl isophthalic acid, 2,3-thiadiazoles-5-ethyl formate (NK-F003) and N-(5-methyl-1,3-thiazole-2-yl)-4-methyl-1,2,3-thiadiazole (SZG-7) directly antiviral activity are very poor, show resistance (table 2) to TMV but these compounds all have in various degree induced activity to tobacco.

Table 2 different concns compound is to the effect of inducing of botrytis cinerea pers

ND: undetermined

The medicament BTH that uses among the present invention, TDL, SZG-7 and NK-F001, NK-F002, NK-F003 mixes use or these medicaments and DL-beta-aminobutyric acid with any two or three, virazole, Ningnanmycin, antofine, No. 1, virus star, No. 2, virus star and XY-13, XY-30 etc. are combined into 2 yuan or 3 yuan of mixtures with arbitrary combination all can obtain the same effect of inducing, the binary mixed ratio of these medicaments is in 1%: 99% to 99%: 1% scope, and the ternary mixed ratio of these medicaments comprises that the ratio of any medicament in mixture all can obtain the effect of inducing of the foregoing description 2 and embodiment 6 and embodiment 7 between 1%: 99%.The formulation that these medicaments can be processed comprises wettable powder, sustained release dosage, aqua, pulvis, micro-capsule suspension, can disperse dense dose, seed treatment emulsion, aqueous emulsion, big granula, granule, microemulsion, oil-suspending agent, finish, the seed with coated pesticidal, suspension concentrates, suspended emulsion agent, water-soluble granule, soluble thick agent, water-dispersible granules etc., and each formulation all can obtain the above-mentioned effect of inducing.

Embodiment 2

Utilize NK-F001, NK-F002, the activity change of phenylalanine ammonia lyase (PAL) in the tobacco body behind NK-F003 and the SZG-7 processing evoking tobacco challenge inoculation TMV

The tobacco leaf that newly grow and that inoculate TMV of learning from else's experience after above-claimed cpd is induced is to be measured, add 3 milliliter of 0.2 mol borate buffer solution (pH8.8, the mercaptoethanol that contains 0.005 mol, 0.001 the EDTA of mol) and the polyvinylpyrrolidone of about example weight 1/10 (PVP), homogenate in ice bath.10000 rev/mins, 4 degree centrifugal 20 minutes down, supernatant liquor is made enzyme activity determination.The mensuration system is by the borate buffer solution of 2 milliliter of 0.2 mol pH8.8.1 milliliter of 0.02 mol L-phenylalanine and 0.8 milliliter of enzyme liquid are formed.After measuring OD290,, measure the OD290 value again, contrast and be 0.8 milliliter of 3 milliliters of enzyme-added liquid of borate buffer solution in 37 degree water-baths 1 hour.The mensuration of protein content adopts Cowes light blue G250 method, is standard protein with BSA.After with medicament was handled the immunity system of evoking tobacco, each is handled the PAL specific activity of enzyme and all is significantly higher than contrast (table 3), and was consistent with the result of biological assay.

Table 3 medicament is induced and is handled the active variation with challenge inoculation back tobacco leaf PAL

Medicament BTH, TDL, SZG-7 and the NK-F001 that uses among the present invention, NK-F002, NK-F003, DL-beta-aminobutyric acid, virazole, Ningnanmycin, antofine, No. 1, viral star, No. 2, viral star and XY-13, XY-30 etc. are combined into 2 yuan or 3 yuan of mixtures with arbitrary combination all can well be induced effect, the binary mixed ratio of these medicaments is in 1%: 99% to 99%: 1% scope, and the ternary mixed ratio of these medicaments comprises that the ratio of any medicament in mixture all can well be induced effect between 1%: 99%.The formulation that these medicaments can be processed comprises wettable powder, sustained release dosage, aqua, pulvis, micro-capsule suspension, can disperse dense dose, seed treatment emulsion, aqueous emulsion, big granula, granule, microemulsion, oil-suspending agent, finish, the seed with coated pesticidal, suspension concentrates, suspended emulsion agent, water-soluble granule, soluble thick agent, water-dispersible granules etc., and each formulation all can obtain the effect of inducing of the foregoing description 1.

Utilize similar method can determine single agent of above-mentioned medicament BTH, TDL, SZG-7 and NK-F001, NK-F002, NK-F003, DL-beta-aminobutyric acid, virazole, Ningnanmycin, antofine, No. 1, viral star, No. 2, viral star and XY-13, XY-30 etc. and be combined into 2 yuan or 3 yuan of mixtures to induce food crop and cash crop and gardening plant to produce resistance to viral diseases of plants with arbitrary combination.

Wherein main crop comprises that cereal crop (comprise paddy, wheat, barley, oat, corn, millet, Chinese sorghum etc.), tuber crops (comprise sweet potato, potato, cassava etc.), legume crop (comprises soybean, broad bean, pea, mung bean, red bean etc.) and fibre crops (cotton, crudefiber crop, silkworm and mulberry etc.), oil crops (peanut, rape, sesame, soybean, Sunflower Receptacle etc.), sugar crop (beet, sugarcane etc.), beverage crops (tealeaves, coffee, cocoa etc.), hobby crop (tobacco leaf etc.), medicinal crop (genseng, the bulb of fritillary etc.), tropical crops (rubber, coconut, oil palm, sisal hemp etc.) food crop and fruit such as, flowers, oil plant, sugar material and cotton, fiber crops, tea, tobacco, cash crop and plantation melons such as Chinese medicinal materials, really, tea, silkworm and mulberry, vegetables (containing various wild vegetable etc.), bamboo shoots, flowers and ornamental plant, hops, medicinal material, pepper, garden crop such as tobacco (flue-cured tobaccos such as seedling and other garden crops, air-curing of tobacco leaves, suncured tabacco), vegetables, (tomato, capsicum, radish, cucumber, Chinese cabbage, celery, hot pickled mustard tube, beet, rape, green onion, garlic etc.), melon (watermelon, muskmelon, hami melon, pawpaw etc.), beans (soybean, broad bean, pea-pods etc.), potato, wheat, corn, paddy rice, peanut, fruit tree, (apple, banana, citrus, peach, papaya), flowers (as orchid), potted landscape etc.; Virus disease comprises tobacco mosaic virus disease, various melon virus diseases, various solanberry viroid diseases, the beans virus disease, the Cruciferae virus disease, the grain and oil crop virus disease, cotton virus disease and various fruit tree virus diseases etc. wherein endanger serious mainly containing: tobacco virus, peppery pimento virus disease, tomato virus disease, Chinese cabbage virus disease, the paddy rice virus disease comprises rice dwarf virus disease, yellow dwart, stripe virus disease, the fern leaf of tomato viral disease, sick and the tobacco veinal necrosis virus disease of chilli pepper mosaic virus, the corn short mosaic disease, cauliflower mosaic virus, the oranges and tangerines virus disease, cymbidium mosaic virus, cybidium ring spot virus etc.

Claims (2)

1.4-methyl isophthalic acid, 2, the purposes of 3-thiadiazoles-5-sodium formiate in preparation evoking tobacco resisting tobacco mosaic virus medicament.

2.4-methyl isophthalic acid, 2, the purposes of 3-thiadiazoles-5-ethyl formate in preparation evoking tobacco resisting tobacco mosaic virus medicament.