CN104396973B - The purposes of trichothecin in anti-tobacco disease fungal pathogens - Google Patents
The purposes of trichothecin in anti-tobacco disease fungal pathogens Download PDFInfo
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- CN104396973B CN104396973B CN201410604123.6A CN201410604123A CN104396973B CN 104396973 B CN104396973 B CN 104396973B CN 201410604123 A CN201410604123 A CN 201410604123A CN 104396973 B CN104396973 B CN 104396973B
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Abstract
The invention discloses a kind of method suppressing tobacco disease fungal pathogens to grow, the method comprises and being contacted with tobacco disease fungal pathogens by trichothecin.The invention also discloses a kind of method of preventing and treating the fungal disease of tobacco, the method comprises trichothecin is applied to tobacco plant.In addition, the invention also discloses the purposes of trichothecin in the fungal disease of anti-tobacco.The present invention can prevent and treat the fungal disease (particularly Alternaria alternate and/or black shank) of tobacco effectively, significant with raising tobacco production aspect to control tobacco diseases.
Description
Technical field
The present invention relates to the purposes of trichothecin in anti-tobacco disease fungal pathogens, particularly, relate to a kind of trichothecin that utilizes and suppress the growth of tobacco disease fungal pathogens, the method for preventing and treating the fungal disease of tobacco and the purposes of trichothecin in the fungal disease of anti-tobacco.
Background technology
Tobacco disease fungal pathogens is one of main chief culprit hindering tobacco production.Wherein, Alternaria alternate is one of Major Diseases on China's tobacco leaf production.There is the generation of this disease in Ge Chanyan district of China, occurs serious with Shandong, Henan, Anhui, Heilungkiang, Jilin and other places; Next is Sichuan, Yunnan, Guizhou, Liaoning, Deng Yan district, Shaanxi.In addition, the Turkish tobaccos producing region in Guangdong, the Dark sun-cured producing region in Zhejiang are endangered also heavier.
Black shank is one of destructive disease of most on tobacco leaf production, also known as tobacco epidemic disease.Each main Chan Yan district of China all has and occurs in various degree, and wherein Anhui, Shandong, Henan Province are historical grave illness district; The Main Tobacco-growing Regions In Souths such as Yunnan, Guizhou, Sichuan, Hunan, Guangdong, Guangxi, Fujian occur also quite general, and many mixing with tobacco bacterial wilt occurs, therefore endanger even more serious.China is owing to taking to change kind of disease-resistant variety for main comprehensive preventive health measures, and the state of an illness is controlled substantially.But in recent years due to some local continuous cropping vega enlarged areas, the continuous cropping time limit extends or changes winter cigarette is spring cigarette, and this disease causes harm the gesture of obvious rising.
Because varieties of plant structure is single in tobacco leaf production, tobacco bred resistance recession, takes the appearance causing " three R " problem as the leading factor with chemical pesticide control, the pesticide resistance of damage by disease and insect strengthens, cause the harm of damage by disease and insect to tobacco to have the trend increased the weight of gradually, constrain the sustainable development of tobacco agriculture.Therefore, meet environmental protection, concern that the biopesticide of the sound and sustained development theory more and more causes people in tobacco disease Pest control, especially the application of microorganism and active secondary metabolite thereof, there is the new tool that broad spectrum activity and environmentally safe advantage have become plant pest management, show wide application prospect, therefore, seek and more have great importance for the medicament preventing and treating tobacco diseases.
Summary of the invention
The object of the invention is to overcome the deficiencies in the prior art, a kind of method of anti-tobacco disease fungal pathogens is newly provided.
To achieve these goals, the present inventor has carried out great many of experiments, found that, trichothecin has excellent effect to suppressing infecting of tobacco disease fungal pathogens.Therefore, first aspect, the invention provides a kind of method suppressing tobacco disease fungal pathogens to grow, and the method comprises and being contacted with tobacco disease fungal pathogens by trichothecin.
Second aspect, the invention provides a kind of method of preventing and treating the fungal disease of tobacco, and the method comprises trichothecin is applied to tobacco plant.
The third aspect, the invention provides the purposes of trichothecin in the fungal disease of anti-tobacco.
By technique scheme, the present invention can prevent and treat the fungal disease (particularly Alternaria alternate and/or black shank) of tobacco effectively, significant with raising tobacco production aspect to control tobacco diseases.
Other features and advantages of the present invention are described in detail in embodiment part subsequently.
Embodiment
Below the specific embodiment of the present invention is described in detail.Should be understood that, embodiment described herein, only for instruction and explanation of the present invention, is not limited to the present invention.
In the present invention, when not doing contrary explanation, the term of use " suppresses the growth of tobacco disease fungal pathogens " and refers to the Growth and reproduction of tobacco disease fungal pathogens suppressed, and concrete manifestation is mycelia or spore death after a period of time.The term " control " used in the present invention means to prevent and/or treat." trichothecin (4 β-the Crotonyloxy-12 used in the present invention, 13-epoxytrichothec-9-en-8-one, Trichothecin) " be the bioactivator with sequiterpene (alkene) or sesquiterpenoid chemical constitution, can from Fusarium (Fusarium), genus (Cylindrocarpon) embraced by post, Myrothecium (Myrothecium), Stachybotrys (Stachybotrys), the mould genus of knurl spore (Yerrucaria), Trichothecium (Trichothecium), extract in the tunning of the imperfect fungus fungies such as cephalo mould (Cephalosporium) and trichoderma (Trichoderma) and obtain, its structure is such as formula shown in (I).
The method of suppression tobacco disease fungal pathogens provided by the invention growth comprises and being contacted with tobacco disease fungal pathogens by trichothecin.
According to the present invention, to the not special requirement of the consumption of described trichothecin, but relative to 10
5individual tobacco disease fungal pathogens, the consumption of described trichothecin is preferably 15-170 μ g, is more preferably 67 μ g, wherein, the quantity of the spore that the quantity of tobacco disease fungal pathogens produces in tobacco disease fungal pathogens.
According to the present invention, described tobacco disease fungal pathogens can for the common disease fungus causing tobacco fungal infection, under preferable case, described tobacco disease fungal pathogens is tobacco brown spot pathogen (Alternariaalternata) and/or tobacco black shank bacterium (Phytophthoranicotianae).
The method of fungal disease of control tobacco provided by the invention comprises trichothecin is applied to tobacco plant.
According to the present invention, described trichothecin can method of application routinely be used tobacco plant, preferably, described trichothecin is used as a solution and the concentration of single-ended sp cellulose solution is 50-500 μ g/mL, be more preferably 100-200 μ g/mL, more preferably 200 μ g/mL.
According to the present invention, to the amount of application of described trichothecin, there is no particular limitation, can be 10-30mg/ strain/time, but preferably, the amount of application of described trichothecin be 20-30mg/ strain/time, be more preferably 25mg/ strain/time (with 8-9 leaf phase tobacco plant for benchmark).Normally, as required, frequency of administration is for use once every 5-7 days.
According to the present invention, in order to prevent and treat the fungal disease of tobacco better, the mode used is preferably spray application.
According to the present invention, described fungal disease is preferably Alternaria alternate and/or black shank.
Present invention also offers the purposes of trichothecin in the fungal disease of anti-tobacco.Preferably, described fungal disease is Alternaria alternate and/or black shank.
Below will be described the present invention by embodiment.
In following examples, not specified experimental implementation is carried out all under normal conditions.Tobacco brown spot pathogen is numbered purchased from ATCC, ATCC
96153
tM; Tobacco black shank bacterium is numbered purchased from ATCC, ATCC
mYA-4101
tM; Experiment is with tobacco the tobacco being planted in Jie Tou town, Tengchong County, Yunnan Province; The trichothecin reference literature that uses " Yang Peiwen etc., the separation and purification of the trichothecin that dark two spore bacterium AGR0073 produces, structure elucidation and anti-fungal activity of plant pathogenic thereof, southwestern agriculture journal, 2014,27 (1): 142-146 " is obtained.
Embodiment 1
The present embodiment is used for the effect adopting trichothecin to suppress the growth of tobacco disease fungal pathogens is described.
The spore getting tobacco brown spot pathogen and tobacco black shank bacterium is respectively made into the spore suspension of debita spissitudo (under 10 × 10 low power lens, each visual field 30-40 spore).Trichothecin is mixed with certain density aqueous acetone solution, then mixes with spore suspension equal-volume, get and be one after another drop ofly added on concave slide, each process 3 repeats, and at 28 DEG C, moisturizing checks the sprouting situation of contrast spore after cultivating 18h.After the germination rate of contrast spore reaches 85%, check the spore germination rate of all process, spore length half person is greater than for sprouting with spore germ tube length, find out trichothecin concentration when inhibition of germination is 100%, be minimum inhibitory concentration (MinimumInhibitoryConcentration, MIC).Result shows, and trichothecin is 6.5 μ g/mL to the MIC of tobacco brown spot pathogen, is 7 μ g/mL to the MIC of tobacco black shank bacterium.Minimum inhibitory concentration is lower, illustrate that bacteriostatic activity is stronger, and the MIC value of trichothecin to tobacco brown spot pathogen and tobacco black shank bacterium is all less than 10 μ g/mL, illustrates that the inhibition of trichothecin to tobacco brown spot pathogen and tobacco black shank bacterium is all stronger.
Embodiment 2
The present embodiment is used for the effect adopting trichothecin to suppress tobacco disease fungal pathogens mycelial growth is described.
Adopt plate growth rate method to measure the inhibition of trichothecin to tobacco brown spot pathogen and tobacco black shank bacterium mycelial growth, step is as follows:
Be the single-ended sp cellulose solution of 500 μ g/ml, 250 μ g/ml, 125 μ g/ml, 62.5 μ g/ml, 31.25 μ g/ml, 15.625 μ g/ml, 7.8125 μ g/ml, 3.90625 μ g/ml respectively with sterile water compound concentration, compare (CK) with sterile water.
Each solution of 10mL dilution is poured in the medium of 8 bottles of sterilizings respectively and (often bottledly has 90mL medium, liquid is respectively by dilution 10 times), after mixing, medium is poured into the culture dish of diameter 9cm, make the pastille culture plate of series concentration gradient, often kind of concentration 5 repeats, stand-by after condensation.Punch cultivating on the medium covering with mycelia with the sterilizing stainless steel card punch of diameter 5mm, sterile working.Mycelia agar block is inoculated in the medium central authorities of condensation, 28 DEG C of cultivations.
Wait contrasting growth close to when covering with culture dish, measuring colony diameter by right-angled intersection method (mean value getting bacterium colony radiation growth major diameter and minor axis diameter deducts primordial hypha block diameter), calculating the mycelial growth inhibition rate of medicament.Computing formula is as follows:
Mycelial growth inhibition rate (%)=(CK bacterium colony average diameter-process bacterium colony average diameter)/CK bacterium colony average diameter × 100%
Mycelial growth inhibition rate value (y) according to each concentration values (x) of trichothecin and correspondence does regression analysis, obtain virulence regression equation (y=a+bx) and correlation coefficient (r), go out according to regression equation calculation the EC that trichothecin suppresses germ mycelial growth
50, EC
90with 95% confidence limit, shown in table 1 specific as follows and 2.
Table 1 trichothecin is to tobacco brown spot pathogen mycelial growth inhibitory action and statistics
Table 2 trichothecin is to the growth inhibition effect of black shank mycelia and statistics
As can be seen from Table 1 and Table 2, the inhibitory action of trichothecin to the mycelial growth of tobacco disease fungal pathogens is stronger.
Embodiment 3
The present embodiment is used for illustrating and adopts trichothecin to infect the mensuration of greenhouse pot culture control efficiency to tobacco disease fungal pathogens.
The consistent health tobacco plant of 8-9 leaf phase growing way is selected to carry out the mensuration of protective effect and therapeutic action.
Protective effect: spray trichothecin (spraying with the concentration of 200 μ g/mL), after 24 hours, spray-on process inoculum density is 3 × 10
5the spore suspension of the tobacco disease fungal pathogens (tobacco brown spot pathogen or tobacco black shank bacterium) of individual/mL, if 1% glucose solution is blank, the amount of spraying of three is 50mL/ strain.Inoculation is placed on moisturizing in 28 DEG C of incubators and cultivates 6d.
Therapeutic action: spray-on process inoculum density is 3 × 10
5the spore suspension of the tobacco disease fungal pathogens (tobacco brown spot pathogen or tobacco black shank bacterium) of individual/mL, if 1% glucose solution is blank, is then placed in 28 DEG C of incubator moisturizings and cultivates 24h.Spray trichothecin (spraying with the concentration of 200 μ g/mL), the amount of spraying of three is 50mL/ strain.5d is cultivated with being placed on moisturizing in 28 DEG C of incubators.
7d " Invest, Then Investigate " result of the test: observe each process blade incidence, statistics disease index, and calculate disease index and control efficiency.
Investigation grade scale:
0 grade: complete stool is anosis; 1 grade: scab number less than 2;
3 grades: scab number 3-5; 5 grades: scab number 6-8;
7 grades: scab number more than 9; 9 grades: scab gathers, and blade is seriously withered.
Disease index=∑ (the sick number of sheets × relative level numerical value at different levels)/(investigating total leaf sum × 9) × 100%
Control efficiency (%)=(control treatment disease index-dispenser process disease index)/contrast disease index × 100%
Trichothecin to tobacco disease fungal pathogens to prevent and treat test result as follows:
Protective effect to Alternaria alternate: repeating average control efficiency for 4 times is 98.12%;
Therapeutic action to Alternaria alternate: repeating average control efficiency for 4 times is 85.33%;
Protective effect to black shank: repeating average control efficiency for 4 times is 96.28%;
Therapeutic action to black shank: repeating average control efficiency for 4 times is 79.61%.
Can find out, trichothecin has control efficiency well to tobacco disease fungal pathogens.
More than describe the preferred embodiment of the present invention in detail; but the present invention is not limited to the detail in above-mentioned embodiment, within the scope of technical conceive of the present invention; can carry out multiple simple variant to technical scheme of the present invention, these simple variant all belong to protection scope of the present invention.
It should be noted that in addition, each concrete technical characteristic described in above-mentioned embodiment, in reconcilable situation, can be combined by any suitable mode, in order to avoid unnecessary repetition, the present invention illustrates no longer separately to various possible combination.
In addition, also can be combined between various different embodiment of the present invention, as long as it is without prejudice to thought of the present invention, it should be considered as content disclosed in this invention equally.
Claims (7)
1. the method suppressing tobacco disease fungal pathogens to grow, the method comprises and being contacted with tobacco disease fungal pathogens by trichothecin, and described tobacco disease fungal pathogens is tobacco brown spot pathogen (Alternariaalternata) and/or tobacco black shank bacterium (Phytophthoranicotianae).
2. method according to claim 1, wherein, the quantity of the spore produced in tobacco disease fungal pathogens, relative to 10
5individual tobacco disease fungal pathogens, the consumption of described trichothecin is 15-170 μ g.
3. prevent and treat a method for the fungal disease of tobacco, the method comprises trichothecin is applied to tobacco plant, and wherein, described fungal disease is Alternaria alternate and/or black shank.
4. method according to claim 3, wherein, described trichothecin is used as a solution and the concentration of single-ended sp cellulose solution is 50-500 μ g/mL.
5. the method according to claim 3 or 4, wherein, the amount of application of described trichothecin be 10-30mg/ strain/time.
6. method according to claim 3, wherein, the mode used is spray application.
7. the purposes of trichothecin in the fungal disease of anti-tobacco, wherein, described fungal disease is Alternaria alternate and/or black shank.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1510573A1 (en) * | 2003-09-01 | 2005-03-02 | Austria Wirtschaftsservice Gesellschaft mit beschränkter Haftung | Method for detoxification of mycotoxins |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| EP1510573A1 (en) * | 2003-09-01 | 2005-03-02 | Austria Wirtschaftsservice Gesellschaft mit beschränkter Haftung | Method for detoxification of mycotoxins |
Non-Patent Citations (1)
| Title |
|---|
| 暗双孢菌AGR0073产生的单端孢菌素的分离纯化、结构解析及其抗植物病原真菌活性;杨佩文 等;《西南农业学报》;20140228;第27卷(第1期);第142-146页 * |
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