CN105594715A - Novel application of TOR (Target of Rapamycin) protein inhibitor to inhibition of late blight bacteria of solanaceous plants - Google Patents
Novel application of TOR (Target of Rapamycin) protein inhibitor to inhibition of late blight bacteria of solanaceous plants Download PDFInfo
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- CN105594715A CN105594715A CN201610171677.0A CN201610171677A CN105594715A CN 105594715 A CN105594715 A CN 105594715A CN 201610171677 A CN201610171677 A CN 201610171677A CN 105594715 A CN105594715 A CN 105594715A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/90—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having two or more relevant hetero rings, condensed among themselves or with a common carbocyclic ring system
Abstract
The invention discloses novel application of a TOR (Target of Rapamycin) protein inhibitor to inhibition of late blight bacteria of solanaceous plants. The TOR protein inhibitor is Rapamycin, KU-0063794, Torin 1, AZD8055, a mixture of Rapamycin and KU-0063794, a mixture of Rapamycin and Torin 1 or a mixture of Rapamycin and AZD8055; the inhibition rate of the late blight bacteria of the solanaceous plants can reach 90%-100%, and a very significant foundation is provided for researches of new chemicals for preventing and treating late blight; and the TOR protein inhibitor has a wide prospect when applied to a novel bactericide, has good market prospects, and provides a new selection for preventing and treating the late blight.
Description
Technical field
The present invention relates to the new purposes of TOR protein inhibitor, relate in particular to TOR protein inhibitor and suppress eggplantThe new purposes of section's plant late disease bacteria.
Background technology
Potato is global the fourth-largest cereal crops, is only second to wheat, paddy rice and corn. And by phytophthora infestans(Phytophthorainfestans) late blight of potato causing, is a kind of fungal disease, can cause horseBell potato cauline leaf is dead and stem tuber is rotten, bring crushing harm can to the production of potato, even can causePotato Severe Reduction even has no harvest. Phytophthora infestans also can cause the plant of Solanaceae such as tomato, capsicum generation late blightSick. Late blight in form by several parts such as mycelia, sporangium, sporangiophore, egg spore, zoosporesComposition. Hyphae colorless, without every, multinuclear, 20～23 DEG C of growth thermophilics, insert in plant cell with haustorium,By drawing plant cell nutrition, thereby expand in a large number numerous.
Current, the method for the prevention and control late blight of potato has chemical prevention and biological control, and main method is chemistryPesticide control. Preventing and treating, chemical agent aspect late blight is numerous, for example: Bordeaux mixture, native copper, oxidationCuprous, zineb, captan, folpet, Bravo, fentinacetate, white urea cyanogen, metalaxyl, white Wheeling.But late blight has all produced resistance to many chemical pesticides at present, and most typical representative is exactly metalaxyl. FirstFrost spirit is born in 1977, and it was unsurpassed to the killing action of late blight at that time, very soon in the whole worldPromote, but within 1979, just having occurred the resistance to the action of a drug. Biological control can overcome that chemical insecticide causesThe resistance to the action of a drug and problem of environmental pollution, utilize the means prevention and control late blight of biological control to have certain entering at presentExhibition. For example: Jiang Jizhi etc. study discovery, the extract of the plants such as onion, garlic, cloves is to late blight sporeSon forms with sprouting fabulous inhibitory action. Wang Shutong research finds that 1% anemarrhena asphodefoides extract is to potato late blightDisease has inhibitory action. Yao Yanpo has reported that the metabolite of Trichoderma can generate late blight of potato spore,Mycelial growth has inhibition to a certain degree. Wait research to find YX paecilomycerol Fermented Condensed thing from heart multitude(YFC) in the time that concentration is 2g/L, can be up to 63.00% to the inhibiting rate of late blight mycelia growth. But,As emerging science, also there are a lot of weak points in biological control, for example: and with chemical bactericide comparison,Stability is lower, operates more complicatedly, yields poorly, and takes effect slow.
TOR (TargetofRapamycin) is very conservative serine/threonine protein kinase in a kind of evolutionEnzyme promotes propagation and the growth of cell in yeast, animal, plant by integrating nutrition and energy signal.TOR signal path in the growth of animal embryo formation, separate living tissue activation, plant roots and leaf, bloom, declineIn the control of old and life cycle etc., all play a part crucial. Research discovery in recent years, TOR also has tuneThe function that save ribosomal biosynthesis, promote translation, metabolism regulates and suppress autophagy. Moving in patternIn thing nematode, fruit bat and mouse, yeast, the disappearance of TOR gene can cause organism embryonic death, saysBright TOR gene is the necessary gene in organism, in the process of growing of biology, is playing the part of central role.Rapamycin (Rapamycin) is a kind of novel macrolide immunosuppressants, is most typical TORProtein inhibitor, is also the representative of TOR albumen first generation inhibitor, it can with the FRB of TOR albumenDomain and FKBP12 combination, suppress TOR protein active. Along with TOR signal path is studied notDeeply disconnected, except thunder handkerchief enzyme element, the inhibitor of TOR albumen also has KU-0063794 at present, Torin1,The two generations inhibitor such as AZD8055. TOR protein inhibitor is mainly used in clinical medical research, pin at presentTo the disease such as tumour, immunosupress of human body, part Study is grown and metabolic pathway for plantRegulatory mechanism etc., but yet there are no by the control that the TOR protein active of Antifungi carries out plant diseaseRelevant report.
Summary of the invention
The object of the invention is provides a kind of TOR protein inhibitor to suppress plant of Solanaceae late blight for above problemThe new purposes of germ.
The technical scheme that adopted is for achieving the above object: a kind of TOR protein inhibitor is planted in inhibition SolanaceaeApplication in thing late disease bacteria.
Described TOR protein inhibitor be Rapamycin, KU-0063794, Torin1, AZD8055,The mixture of mixture, Rapamycin and the Torin1 of Rapamycin and KU-0063794 orThe mixture of Rapamycin and AZD8055.
As preferably, application mode is that described TOR protein inhibitor is prepared as to bactericide.
In technique scheme, in described bactericide, the concentration of Rapamycin is 0.01～3 μ m/L,The concentration of KU-0063794 is 0.1～5 μ m/L, and the concentration of Torin1 is 0.1～10 μ m/L, and AZD8055's is denseDegree is 0.005～0.2 μ m/L.
As preferably, the concentration of described Rapamycin is 0.05～3 μ m/L, described KU-0063794'sConcentration is 0.5～5 μ m/L, and the concentration of described Torin1 is 0.5～10 μ m/L, and the concentration of described AZD8055 is0.01～0.2μm/L。
As preferably, in the mixture of described Rapamycin and KU-0063794, Rapamycin's is denseDegree is 0.05～3 μ m/L, and the concentration of KU-0063794 is 2～5 μ m/L.
As preferably, in the mixture of described Rapamycin and Torin1, being combined as of mixture:The concentration of Rapamycin is 0.1～3 μ m/L, and the concentration of Torin1 is 0.5～10 μ m/L; Or the group of mixtureBe combined into: the concentration of Rapamycin is 0.05～3 μ m/L, the concentration of Torin1 is 2～10 μ m/L.
As preferably, in the mixture of described Rapamycin and AZD8055, being combined as of mixture:The concentration of Rapamycin is 0.1～3 μ m/L, and the concentration of AZD8055 is 0.01～0.2 μ m/L; Or mixtureBe combined as: the concentration of Rapamycin is 0.05～3 μ m/L, and the concentration of AZD8055 is 0.05～0.2 μ m/L.
The invention has the beneficial effects as follows: the present invention will be used for first clinical medical TOR protein inhibitor forIn the study on prevention of plant of Solanaceae late blight, by research find TOR protein inhibitor Rapamycin,KU-0063794, Torin1, AZD8055 and Rapamycin respectively with KU-0063794, Torin1,The mixture of AZD8055 all can suppress plant of Solanaceae late disease bacteria effectively, and inhibiting rate can reach 90-100%,Provide very significant basis for preventing and treating late blight new drug research, the prospect that is applied to new type bactericide is wideWealthy, market prospects are good, provide a kind of new selection for preventing and treating late blight.
Brief description of the drawings
Fig. 1 is for blank and add the 5th, 9,13 days colony growth situations of DMSO solvent control experiment.
Fig. 2 is the 9th day colony growth situation of Rapamycin+KU-0063794 combined treatment late disease bacteria.
Fig. 3 is the 13rd day colony growth situation of Rapamycin+KU-0063794 combined treatment late disease bacteria.
Fig. 4 is the 9th day colony growth situation of Rapamycin+Torin1 combined treatment late disease bacteria.
Fig. 5 is the 13rd day colony growth situation of Rapamycin+Torin1 combined treatment late disease bacteria.
Fig. 6 is the 9th day colony growth situation of Rapamycin+AZD8055 combined treatment late disease bacteria.
Fig. 7 is the 13rd day colony growth situation of Rapamycin+AZD8055 combined treatment late disease bacteria.
Detailed description of the invention
Main agents used in the present invention:
Rapamycin, AZD8055, KU-0063794, Torin1 are all purchased from U.S. SelleckChemicalsCompany.
Rapamycin (Sirolimus, rapamycin): be a kind of specific mTOR inhibitors, catalog number (Cat.No.) S1039.
AZD8055: be the competitive mTOR inhibitors of a kind of novel ATP, catalog number (Cat.No.) S1555.
KU-0063794: act on the dual mTOR inhibitors of mTORC1 and mTORC2, catalog number (Cat.No.) S1226.
Torin1:mTORC1/2 inhibitor, catalog number (Cat.No.) S2827.
Phytophthora infestans (phytophthora infestans) is carried by University Of Chongqing's Life Science College plant molecular breeding laboratorySupply.
Embodiment 1 prepares phytophthora infestans culture medium
Phytophthora infestans culture medium adopts rye solid medium, and concrete configuration method is: 1. that 60g is blackWheat is immersed in 300ml distilled water, and 4 DEG C are soaked 36 hours, avoid rotten. 2. immersion water is poured out to preservationIn refrigerator. 3. in the rye expanding, add 100ml distilled water to smash 1-2min to pieces, its suspension is 50Degree water-bath 3h. 4. use 3-4 layer filtered through gauze suspension, preserve filtrate. 5. immersion water second step being obtainedMix with the filtrate that the 4th step obtains, in mixed liquor, add 20g sucrose, dissolve, then add water to 1000ml,Mix. 6. add 12g agar, autoclaving 121 is spent 20min.
After sterilizing, take out nutrient solution, in the time being cooled to about 50 DEG C, add respectively embodiment 2-4 by experiment combinationThe TOR protein inhibitor combination of middle combination of two: Rapamycin+KU-0063794, Rapamycin+TorinThe combination of 1, Rapamycin+AZD8055 variable concentrations, mixes, and pours in culture dish each culture dish intoIn pour 25mL nutrient solution into, each concentration combination fall 3 flat boards do 3 repetitions. Condensation Ma Ling to be inoculatedPotato late disease bacteria.
Get phytophthora infestans dish, get bacterium piece with 0.7cm rifle head, each dull and stereotyped central authorities inoculate 1 bacterium pieceMake each flat board inoculate the phytophthora infestans bacterium piece of 0.7cm diameter. At 18 DEG C, dark conditionUnder cultivate, within the 5th, 9,13 days, measure colony diameters taking pictures respectively at cultivating, calculate bacteriostasis rate. WithTime, to add the DMSO solvent (dimethyl sulfoxide (DMSO)) of maximum concentration 230 μ l/ml as contrast, do not add DMSOBlank and the flat board colony growth situation of the 5th, 9,13 days that added DMSO as shown in Figure 1, bacteriumSpot diameter is as shown in table 1, and blank is consistent, visible with the dull and stereotyped colony growth situation that has added DMSODMSO there is no inhibitory action for the growth of phytophthora infestans.
Table 1 blank and the 5th day, 9 days, 13 days colony diameter statistical forms of DMSO solvent control experiment
The embodiment 2TOR protein inhibitor Rapamycin+KU-0063794 combined treatment late blight of potatoBacterium
The concentration gradient of setting Rapamycin is respectively: 0,0.05,0.1,0.5,1.0,3.0 μ m/L,The concentration gradient of KU-0063794 is: 0,0.5,1.0,2.0,5.0 μ m/L, each concentration combination repeats 3Inferior. The variable concentrations combination bacteriostasis rate of the 5th, 9,13 days is respectively in the (bacterium of the 9th, 13 days shown in table 2～4The long figure that is born is shown in shown in Fig. 2,3):
The 5th day bacteriostasis rate of table 2Rapamycin+KU-0063794 variable concentrations combined treatment phytophthora infestans
The 9th day bacteriostasis rate of table 3Rapamycin+KU-0063794 variable concentrations combined treatment phytophthora infestans
The 13rd day bacteriostasis rate of table 4Rapamycin+KU-0063794 variable concentrations combined treatment phytophthora infestans
From above result, the combination of Rapamycin and KU-0063794, inhibition increases with concentrationAnd increase progressively, and cooperative effect is obvious, and two medicine treatment effects are significantly better than single medicine processing. KU-0063794 is denseDegree is that 2 and 5 μ m/L and Rapamycin concentration are 0.05,0.1,0.5,1.0, the situation of 3.0 μ m/L combinationsUnder, the effect that suppresses the growth of late blight mycelia is best, and Phytophthora infestans inhibiting rate can reach 90-100%.
Embodiment 3TOR protein inhibitor Rapamycin+Torin1 combined treatment phytophthora infestans
The concentration gradient of setting Rapamycin is respectively: 0.05,0.1,0.5,1.0,3.0 μ m/L, Torin1Concentration gradient be: 0.5,2.0,5.0,10.0 μ m/L, each concentration combination repeats 3 times. Variable concentrations groupClose the 5th, 9,13 days bacteriostasis rate respectively in shown in table 5～7 (the colony growth figure of the 9th, 13 days see Fig. 4,Shown in 5):
The 5th day bacteriostasis rate of table 5Rapamycin+Torin1 variable concentrations combined treatment phytophthora infestans
The 9th day bacteriostasis rate of table 6Rapamycin+Torin1 variable concentrations combined treatment phytophthora infestans
The 13rd day bacteriostasis rate of table 7Rapamycin+Torin1 variable concentrations combined treatment phytophthora infestans
From above result, the combination of Rapamycin and Torin1, inhibition increases and passs with concentrationIncrease, and cooperative effect is obvious, two medicine treatment effects are significantly better than single medicine processing. Torin1 concentration is 0.5 μ m/LWith Rapamycin concentration be 0.1,0.5,1.0,3.0 μ m/L combinations, and Torin1 concentration is 2,5,10 μ m/L and Rapamycin concentration are 0.05,0.1,0.5,1.0, and 3.0 μ m/L combinations, suppress late blightThe effect of mycelial growth is best, and inhibiting rate can reach 90-100%.
Embodiment 4TOR protein inhibitor Rapamycin+AZD8055 combined treatment phytophthora infestans
The concentration gradient of setting Rapamycin is respectively: 0.05,0.1,0.5,1.0,3.0 μ m/L, AZD8055Concentration gradient be: 0.01,0.05,0.1,0.2 μ m/L, by the Rapamycin of each concentration gradient andAZD8055 combination, each concentration combination in triplicate. The variable concentrations combination bacteriostasis rate of the 5th, 9,13 daysRespectively in (the colony growth figure of the 9th, 13 days is shown in shown in Fig. 6,7) shown in table 8～10:
The 5th day bacteriostasis rate of table 8Rapamycin+AZD8055 variable concentrations combined treatment phytophthora infestans
The 9th day bacteriostasis rate of table 9Rapamycin+AZD8055 variable concentrations combined treatment phytophthora infestans
The 13rd day bacteriostasis rate of table 10Rapamycin+AZD8055 variable concentrations combined treatment phytophthora infestans
From above result, the combination of Rapamycin and AZD8055, inhibition is with concentration increaseIncrease progressively, and cooperative effect is obvious, two medicine treatment effects are significantly better than single medicine processing. AZD8055 concentration is0.01 μ m/L and Rapamycin concentration are 0.1,0.5,1.0,3.0 μ m/L combinations, and AZD8055 is denseDegree is that 0.05,0.1,0.2 μ m/L and Rapamycin concentration are 0.05,0.1,0.5,1.0,3.0 μ m/L groupsClose, the effect that suppresses the growth of late blight mycelia is best, and inhibiting rate can reach 90-100%.
Claims (8)
1. a TOR protein inhibitor is in the application suppressing in plant of Solanaceae late disease bacteria.
2. application according to claim 1, is characterized in that: described TOR protein inhibitor isRapamycin, KU-0063794, Torin1, AZD8055, Rapamycin and KU-0063794's is mixedThe mixture of the mixture of compound, Rapamycin and Torin1 or Rapamycin and AZD8055.
3. application according to claim 2, is characterized in that: application mode is by described TORProtein inhibitor is prepared as bactericide.
4. application according to claim 3, is characterized in that: Rapamycin in described bactericideConcentration is 0.01～3 μ m/L, and the concentration of KU-0063794 is 0.1～5 μ m/L, and the concentration of Torin1 is 0.1～10μ m/L, the concentration of AZD8055 is 0.005～0.2 μ m/L.
5. application according to claim 4, is characterized in that: the concentration of described Rapamycin is0.05～3 μ m/L, the concentration of described KU-0063794 is 0.5～5 μ m/L, the concentration of described Torin1 is0.5～10 μ m/L, the concentration of described AZD8055 is 0.01～0.2 μ m/L.
6. application according to claim 5, is characterized in that: described Rapamycin and KU-0063794Mixture in, the concentration of Rapamycin is 0.05～3 μ m/L, the concentration of KU-0063794 is 2～5 μ m/L.
7. application according to claim 5, is characterized in that: described Rapamycin and Torin1Mixture in, being combined as of mixture: the concentration of Rapamycin is 0.1～3 μ m/L, the concentration of Torin1Be 0.5～10 μ m/L; Or the concentration of being combined as of mixture: Rapamycin is 0.05～3 μ m/L, Torin1 concentration is 2～10 μ m/L.
8. application according to claim 5, is characterized in that: described Rapamycin and AZD8055Mixture in, being combined as of mixture: the concentration of Rapamycin is 0.1～3 μ m/L, AZD8055'sConcentration is 0.01～0.2 μ m/L; Or the concentration of being combined as of mixture: Rapamycin is 0.05～3 μ m/L,The concentration of AZD8055 is 0.05～0.2 μ m/L.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110468151A (en) * | 2019-09-04 | 2019-11-19 | 中国农业科学院都市农业研究所 | Fusarium oxysporum TOR gene RNAi carrier and its method for combining prevention and treatment dry rot of potato and wilt disease with salicylic acid |
| CN111436337A (en) * | 2020-03-04 | 2020-07-24 | 沈阳农业大学 | Application of rapamycin target protein activator MHY1485 in promotion of tomato seed germination and seedling growth and development |
| CN112021335A (en) * | 2020-09-10 | 2020-12-04 | 重庆大学 | Method for preventing and treating plant oomycete diseases by combining biological pesticide and chemical pesticide |
| CN113957081A (en) * | 2021-10-25 | 2022-01-21 | 沈阳农业大学 | Gene for regulating and controlling growth and development of tomato epidermal hair and application thereof |
| CN115005220A (en) * | 2022-05-31 | 2022-09-06 | 中国热带农业科学院环境与植物保护研究所 | Application of rapamycin in inhibiting colletotrichum gloeosporioides or fusarium verticillioides |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110468151A (en) * | 2019-09-04 | 2019-11-19 | 中国农业科学院都市农业研究所 | Fusarium oxysporum TOR gene RNAi carrier and its method for combining prevention and treatment dry rot of potato and wilt disease with salicylic acid |
| CN111436337A (en) * | 2020-03-04 | 2020-07-24 | 沈阳农业大学 | Application of rapamycin target protein activator MHY1485 in promotion of tomato seed germination and seedling growth and development |
| CN111436337B (en) * | 2020-03-04 | 2021-10-22 | 沈阳农业大学 | Application of rapamycin target protein activator MHY1485 in promotion of tomato seed germination and seedling growth and development |
| CN112021335A (en) * | 2020-09-10 | 2020-12-04 | 重庆大学 | Method for preventing and treating plant oomycete diseases by combining biological pesticide and chemical pesticide |
| CN113957081A (en) * | 2021-10-25 | 2022-01-21 | 沈阳农业大学 | Gene for regulating and controlling growth and development of tomato epidermal hair and application thereof |
| CN113957081B (en) * | 2021-10-25 | 2023-01-24 | 沈阳农业大学 | Gene for regulating and controlling growth and development of tomato epidermal hair and application thereof |
| CN115005220A (en) * | 2022-05-31 | 2022-09-06 | 中国热带农业科学院环境与植物保护研究所 | Application of rapamycin in inhibiting colletotrichum gloeosporioides or fusarium verticillioides |
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