CN103053624B - Method for control of phytophthora blight of pepper by mixed application of trichoderma preparation and fungicides - Google Patents
Method for control of phytophthora blight of pepper by mixed application of trichoderma preparation and fungicides Download PDFInfo
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Abstract
The present invention discloses a method for control of phytophthora blight of pepper by mixed application of trichoderma preparation and fungicides. The method comprises: seeding after mixing pepper seeds with the trichoderma preparation during seeding, or, applying the trichoderma preparation during pepper seedling transplanting and planting; during pepper cultivation, applying the fungicides before the occurrence of the disease or in early days of the disease; wherein the active strain in the trichoderma preparation is Trichodermaasperellum Thz01, and the preservation number thereof is CGMCC No. 6422; the fungicides are one or both of eugenol and mandipropamid. The trichoderma preparation used in the method of the invention has good control effect on phytophthora blight of pepper in the fields, and can effectively reduce the primary infection source when being used alone; while the additional application of the fungicides can effectively control disease epidemics. According to the method of the invention, the application amount of the fungicides is reduced, so residues of pesticides in the soil and pepper products are decreased, thereby ensuring environmental and product safety.
Description
Technical field
The present invention relates to biological control and the Techniques For Chemical Control field of plant disease, relate in particular to the method for a kind of wooden removing mildew control mixed with bactericide capsicum epidemic disease.
Background technology
Capsicum epidemic disease is commonly called as " dead seedling is sick ", is a kind of soil-borne disease being caused by Phytophthora capsici (Phytophthora capsici), is a kind of destructive disease, can cause serious production loss.Because pathogen produces a large amount of zoospores in the popular phase of disease, can cause rapidly capsicum Large Scale Death in a short time.The conventional anti-control techniques of capsicum epidemic disease has been difficult to effective control, and easily causes ecology, environmental problem.
The application for a patent for invention of publication number CN102415378A discloses a kind of bactericide and application thereof that promotes to grow and prevent and treat capsicum epidemic disease, this bactericide comprises cinnamic acid 0.1～10% according to percentage by weight, emulsifier 10～30%, azone 0.1～10%, alcohol 20～50%, all the other are water.This bactericide can be used for preventing and treating Phytophthora capsici and promotes plant growth, but its used chemicals and plant extracts cost higher, be unfavorable for large-scale application.For many years, chemical control is the major measure of the medium-term and long-term improvement capsicum epidemic disease of production practices, but use in a large number chemical agent on the potential harm of human health, on the pollution of food and environment, on the impact of non-target organism and cause the problems such as the drug-fast accelerated development of pathogen to cause social growing interest, it is the inexorable trend of development that decrement is used chemical pesticide.
In recent years, biological prevention and control agent has been introduced in the aspect that prevents and treats of capsicum epidemic disease, and to capsicum epidemic disease, control is effective to especially wooden mould biological prevention and control agent proof.We,, on the basis of research for many years, have filtered out trichoderma asperellum strain (Trichoderma asperellum) CGMCC6422, and capsicum epidemic disease control efficiency is reached to 70% left and right.Yet, although Trichoderma has saprophytic property, resistance and pathogen is had to lasting antagonism, can long-term surviving, in the medium characteristic of soil, can reduce pathogenic soil inoculum quantity, but the pathogen effect in disease popular period has been shown to limitation.Therefore, screen efficient medicament from the bactericide of current control capsicum epidemic disease, these bactericide are to all unrestraint effects of the mycelia of the mould biological prevention and control agent of wood, spore germination and Sporulation simultaneously, and this is also core content of the present invention.On the basis of wooden removing mildew control, in the popular phase of disease, suitably use a small amount of bactericide and less number of times, can effectively prevent and treat capsicum epidemic disease, thereby reach chemical pesticide decrement, use.
Combine with the decrement bactericide technology report not yet so far of control capsicum epidemic disease of the mould biological prevention and control agent of wood, this technology, on the basis of minimizing bactericide quantity, is effectively controlled the popular of capsicum epidemic disease, thereby has been reduced loss; Meanwhile, reduced the pollution of bactericide and protected ecotope.Accordingly, the present invention proposes meter and has founded combine with the agricultural chemicals new method of Collaborative Control capsicum epidemic disease of a kind of biological control.
Summary of the invention
The invention provides the method for a kind of wooden removing mildew control mixed with bactericide capsicum epidemic disease, in order to solve the simple biological control of capsicum epidemic disease, can not control the problem that disease is popular, decrement is used chemical agent simultaneously.
A method for wooden removing mildew control mixed with bactericide capsicum epidemic disease in pepper planting process, the plantation of capsicum comprises sowing, grows seedlings, transplant planting, cultivate and gather; Described method comprises:
(1) while sowing, sowing after pepper seed is mixed with wooden removing mildew; Or, when Hot Pepper Seedling transplant planting, use wooden removing mildew;
(2) during pepper cultivation, before disease occurs or initial stage of origination use bactericide;
Wherein, the active bacterial strain trichoderma asperellum in described wooden removing mildew (Trichoderma asperellum) Thz01, the preserving number of trichoderma asperellum Thz01 is CGMCC No.6422;
Described bactericide is one or both in eugenol and mandipropamid.
Wooden removing mildew prepared by trichoderma asperellum Thz01 has good preventive effect in field to capsicum epidemic disease, in the disease emergence period, combines use with bactericide, can effectively prevent the generation of disease; The use of energy decrement bactericide simultaneously.Wood removing mildew is generally comprised of active bacterial strain and corresponding auxiliary material, and auxiliary material is not only as carrier, and provides nutrient to strain growth, and the key of epidemic prevention is the selection of active bacterial strain.
In the present invention, described trichoderma asperellum Thz01 preservation, depositary institution: the China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC) that is positioned at Datun Road, Chaoyang District, Beijing City Institute of Microorganism, Academia Sinica, preservation date: on August 13rd, 2012, preserving number: CGMCCNo.6422.According to morphological feature and molecular data analysis, the Classification And Nomenclature of this bacterial strain is trichoderma asperellum (Trichoderma asperellum).
This bacterial strain is to collect from the capsicum rhizosphere soil soil of mountain vegetables garden, Linan morbidity, the biological property of this bacterial strain is: bacterium colony is cultivated at 25 ℃ on PDA solid culture medium, within 3-4 days, expand to 9cm, initial stage white is sparse, mycelia is along media surface sprawl growth, the green Chan Bao of rear formation district, reverse side white, mycelia tool every; Conidium is spherical, subsphaeroidal, single closely colourless, is pistac during gathering, and wall is smooth.Upright, the ampulla shape of bottle, width 3.0-5.6 μ m.The upper conidium of CMD is spherical to sub-spherical, 3.8-5.9 * 2.8-4.8 μ m.Morphological feature meets trichoderma asperellum bacterium feature.According to ITS rDNA(SEQ ID NO.1) and transcription factor 1a(tef1-a) sequence (SEQ ID NO.2) analyzes, the most consistent with the trichoderma asperellum bacterium sequence of pattern in GenBank and confirmation.These confirm that this bacterial strain is trichoderma asperellum (Trichoderma asperellum).
Because wooden removing mildew needs the competence exertion effect of field planting a period of time after using, for this reason, described wooden removing mildew was used the early stage of plantation when Hot Pepper Seedling transplant planting (during sowing or), can be made microorganism wherein be colonizated in soil preferably, to prevent that disease from occurring.
In order further to strengthen the action effect of trichoderma, during pepper cultivation, according to the local capsicum epidemic disease state of an illness, observe and predict information, can within 0.5-1 month before disease occurs, at capsicum rhizosphere, use wooden removing mildew around.Like this, can play the effect of strengthening preventive effect.
While using, the consumption of described wooden removing mildew is preferably 1.5-2g/ strain at every turn, and Microorganism colonization effect is better, and can not cause preparation waste.When using during pepper cultivation, amount of application also can according to local disease, a situation arises considers adjustment.
Described wooden removing mildew can be prepared by the following method: trichoderma asperellum Thz01 spore suspension is inoculated in the wheat that boils sterilizing, in illumination in 12 hours and 12 hours dark alternate culture 15-20 days, makes wooden removing mildew.
Wherein, described trichoderma asperellum Thz01 spore suspension is prepared by the following method: trichoderma asperellum Thz01 is inoculated in PDA medium, at 23-28 ℃, cultivates 5-6 days; After cultivation completes, spore is scraped in aqua sterilisa, preparation obtains trichoderma asperellum Thz01 spore suspension.
The viable bacteria concentration of described trichoderma asperellum Thz01 spore suspension is preferably 10
7-10
8individual/mL.
Described wheat is the matrix that is applicable to trichoderma asperellum Thz01 growth.
In every kilogram of wheat, the consumption of described trichoderma asperellum Thz01 spore suspension is preferably 2-4mL.
Incubation time after trichoderma asperellum Thz01 spore suspension access wheat is preferably 18 days, is convenient to produce in matrix a large amount of spores.
Between culture period, can stir in time wheat, so that fully long upper trichoderma asperellum Thz01 spore on wheat.
Using of described bactericide can be carried out according to operation instruction, generally at disease initial stage of origination, carries out dispenser for the first time, and further dispenser decided depending on the lasting period of chili growth disease in season development and medicament.
During pepper cultivation, according to the local capsicum epidemic disease state of an illness, observe and predict information, before disease occurs or the bactericide of initial stage of origination described in using, can effectively prevent and treat disease.Preferably, can be before disease occurs 1-2 days or the initial stage of origination field bactericide described in using while there is fragmentary scab plant.
Described bactericide can be eugenol; In every mu, the consumption of eugenol active ingredient can be 0.078-0.084g; Be preferably 0.081g.Product (active constituent content is 0.3%) that eugenol can adopt Baoding Ya Da Chemical Co., Ltd. to produce, in every mu, gets 26-28g, is watered 60L, obtains the medicament that active ingredient concentration is 1.3-1.4 μ g/mL, then sprays.Eugenol consumption is very few, does not have control efficiency; Consumption is excessive, can affect trichoderma asperellum Thz01 active, even can kill trichoderma strain.Under this consumption condition, eugenol and wooden removing mildew coupling effect are best.
Described bactericide can be mandipropamid; In every mu, the consumption of mandipropamid active ingredient can be 1.8-2.1g; Be preferably 1.95g.The product (active constituent content is 250g/L) that mandipropamid can adopt Switzerland Xian Zhengda crop protection Co., Ltd to produce; in every mu; get 7.2-8.4g(and be approximately equivalent to 7.2-8.4mL); be watered 30L; obtain the medicament that active ingredient concentration is 60-70 μ g/mL, then spray.Similarly, under this consumption condition, mandipropamid and wooden removing mildew coupling effect are best.
Disease can further be used bactericide according to pepper diseases development after occurring, and strengthens preventive effect.Each amount of application is: eugenol is that active ingredient reaches 0.078-0.084g/ mu, is preferably 0.081g/ mu; Or mandipropamid is that active ingredient reaches 1.8-2.1g/ mu; Be preferably 1.95g/ mu.
Test confirmation, wooden removing mildew and bactericide mix the consumption that use has effectively reduced bactericide, reach the control efficiency identical with chemical control, to wooden removing mildew, are also safe simultaneously.In production, bactericide eugenol (Baoding Ya Da Chemical Co., Ltd.) recommendation amount is 30g/ mu; And wooden removing mildew and eugenol mix use, 27g/ mu eugenol usage amount, with usage amount is identical separately, is equivalent to reduce 3g/ mu, and decrement is used bactericide 10%; Inhibition test demonstration, wooden removing mildew is safe using under this bactericide 30g/ mu condition.Similarly, in production, bactericide mandipropamid (Switzerland Xian Zhengda crop protection Co., Ltd) recommendation amount is 12g/ mu; And wooden removing mildew and mandipropamid mix use, 7.8g/ mu mandipropamid amount of application, with usage amount is identical separately, is equivalent to reduce 4.2g/ mu, and decrement is used bactericide 35%; Inhibition test demonstration, wooden removing mildew is safe using under this bactericide 9.6g/ mu condition.
In the present invention, disease time of origin mainly observes and predicts information judgement according to local meteorological data or the state of an illness.The disease time of the popular disease of capsicum epidemic disease is relevant with season, weather etc., but occurs every year slightly to change period.The time occurring in the plastic greenhouse of Hangzhou as capsicum epidemic disease is in left and right by the end of March, and open country occurs in mid-April.Different regions are because weather conditions are different, and disease time of origin is slightly different, and as In Hangzhou Region of Zhe Jiang Province open country is generally moon mid-April, and Ningbo area generally will be done sth. in advance about 10 days.
The present invention, during pepper planting, in suitable period, in a certain order, adopts the wooden removing mildew of suitable dose and bactericide mixing to make for preventing and treating capsicum epidemic disease.
Adopt the inventive method, there is following beneficial effect:
(1) the wooden removing mildew that prepared by trichoderma asperellum Thz01 has good preventive effect in field to capsicum epidemic disease, and independent wooden removing mildew can effectively reduce primary source of infection.
(2) in the disease emergence period, increase the use of bactericide, can effectively control disease popular.This utilization can decrement bactericide use, the amount of application of bactericide does not have inhibitory action to the growth of Trichoderma, and eugenol reducing application amount 10%, mandipropamid reducing application amount 35%, both effectively controlled the popular of capsicum epidemic disease, reduce again residual in soil and capsicum product of agricultural chemicals, guaranteed the safety of environment and product.
(3) the inventive method is reasonable in design, easy and simple to handle, and input and operating cost are lower, is applicable to large-scale operation.Applying of the method, has facilitation for capsicum health, safety in production and sustainable agriculture development.
Accompanying drawing explanation
Fig. 1 is that wooden removing mildew mixes the rear field of the use capsicum epidemic disease incidence of disease, disease index and control efficiency with the mandipropamid of various dose; Wherein, figure A-C is respectively dispenser the disease incidence of disease, disease index and control efficiency measurement result after 15 days.
Fig. 2 is that wooden removing mildew mixes the rear field of the use capsicum epidemic disease incidence of disease, disease index and control efficiency with the eugenol of various dose; Wherein, figure A-C is respectively dispenser the disease incidence of disease, disease index and control efficiency measurement result after 15 days.
Fig. 3 is two kinds of bactericide inhibitions to phytophthora blight of pepper PC004 and trichoderma asperellum Thz01 respectively; Wherein, figure A and figure C are respectively phytophthora blight of pepper PC004 and cultivate 7 days on the PDA flat board that contains 65 μ g/ml mandipropamids or the former medicine of 1.35 μ g/ml eugenol; Figure B and figure D are respectively trichoderma asperellum Thz01 and cultivate 3 days on the PDA flat board that contains 65 μ g/ml mandipropamids or the former medicine of 1.35 μ g/ml eugenol.
Embodiment
Trichoderma asperellum Thz01 bacterial strain
Trichoderma asperellum (Trichoderma asperellum) Thz01 bacterial strain is to collect from the capsicum rhizosphere soil soil of mountain vegetables garden, Linan morbidity, the biological property of this bacterial strain is: bacterium colony is cultivated at 25 ℃ on PDA solid culture medium, within 3-4 days, expand to 9cm, initial stage white is sparse, mycelia is along media surface sprawl growth, the green Chan Bao of rear formation district, reverse side white, mycelia tool every; Conidium is spherical, subsphaeroidal, single closely colourless, is pistac during gathering, and wall is smooth.Upright, the ampulla shape of bottle, width 3.0-5.6 μ m.The upper conidium of CMD is spherical to sub-spherical, 3.8-5.9 * 2.8-4.8 μ m.Morphological feature meets trichoderma asperellum bacterium feature.According to ITS rDNA(SEQ ID NO.1) and transcription factor 1a(tef1-a) sequence analysis, the most consistent with the trichoderma asperellum bacterium sequence of pattern in GenBank and confirmation.These confirm that this bacterial strain is trichoderma asperellum (Trichoderma asperellum).
The preservation of trichoderma asperellum Thz01 bacterial strain, depositary institution: China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation date: on August 13rd, 2012, preserving number: CGMCCNo.6422.
The preparation of embodiment 1 wooden removing mildew
(1) prepare trichoderma asperellum Thz01 spore suspension: trichoderma asperellum Thz01 is inoculated in PDA medium, at 25 ℃, cultivates 5-6 days; After cultivation completes, spore is scraped in aqua sterilisa, preparation obtains trichoderma asperellum Thz01 spore suspension, and (viable bacteria concentration is 10
8individual/mL).
(2) prepare wooden removing mildew: trichoderma asperellum Thz01 spore suspension is inoculated in the wheat (feed wheat) that boils sterilizing, between culture period, stir in time, at illumination in 12 hours and 12 hours dark alternate cultures after 18 days, on inducing culture, produce a large amount of spores, spore is prepared into wooden removing mildew together with spore.
In every kilogram of wheat, the consumption of trichoderma asperellum Thz01 spore suspension is 3mL.
The screening of embodiment 2 bactericide
Select 13 kinds of bactericide that are usually used in preventing and treating capsicum epidemic disease in producing for screening test, comprising: 80% phosethyl-Al (Zhejiang Jia Hua Chemical Co., Ltd.); 250g/L mandipropamid (Switzerland Xian Zhengda crop protection Co., Ltd); 250g/L Amici reaches (Britain Syngenta Co., Ltd); 72% cymoxanil mancozeb (du pont company); 50% dimethomorph (the biochemical Co., Ltd of Shijiazhuang Yi Nuo); 69% dimethomorph mancozeb (the biochemical Co., Ltd of Ke Lilong); 72% metalaxyl (Kai Lier biochemical corp, Jiangsu); 50% second aluminium MnZn (favorable to the people Chemical Co., Ltd.); 46% Kocide SD (du pont company); 75% tpn (favorable to the people Chemical Co., Ltd.); 0.3% eugenol (Baoding Ya Da Chemical Co., Ltd.); 20% flumorph (Shenyang Kechuang chemicals Co., Ltd); The 66.8% the third gloomy iprovalicarbs (German Bayer Cropscience N.V.).
The screening of medicament is divided into following three processes:
(1) inhibitory action of different bactericide to trichoderma asperellum Thz01 mycelial growth
Adopt mycelial growth rate determination method to measure the inhibitory action of different bactericide to trichoderma asperellum Thz01 mycelial growth, filter out the medicament that bactericide does not suppress trichoderma asperellum Thz01 mycelia.
Method: first measure the concentration range that each Medicaments on Trichoderma viride mycelial growth starts inhibiting rate, then around this concentration 5 series concentration setting up and down.Reagent agent, respectively with being made into certain density mother liquor, is drawn to needed volume according to the concentration of having set, join in triangular flask.Triangular flask contains the PDA medium having melted, conventionally suitable being incorporated as of 45 ℃ of left and right, mixes in the rear culture dish (Φ=90 ㎜) of pouring respectively sterilizing into, take that to add the PDA flat board of equal-volume sterile water be contrast.With card punch (Φ=5 ㎜), cut the Trichoderma bacterium cake of cultivating 2 days on PDA medium, be inoculated in the PDA medium central authorities of pastille, be placed in dark culturing in 25 ℃ of constant incubators, each concentration is processed and is repeated 3 times.Within 3 days, measure afterwards the colony diameter of strains tested on variable concentrations pastille medium, calculate the growth inhibition ratio of each chemicals treatment to bacterium colony expansion with comparing, analyze more different bactericide to the impact for examination germ mycelial growth, calculate the virulence regression equation of medicament.The theoretical value of drug concentration while being 1% according to virulence regression equation calculating inhibiting rate, drug concentration gradient is set on the basis of this theoretical value, under identical training method and condition, to determine and do not suppress the measured value of mycelial growth completely, each concentration is processed and is repeated 3 times.
Result shows, in 13 kinds of bactericide, the poor bactericide of inhibition of trichoderma asperellum Thz01 mycelial growth is had to phosethyl-Al, mandipropamid, cymoxanil mancozeb, dimethomorph, dimethomorph mancozeb, Kocide SD and the third gloomy iprovalicarb, and its virulence approaches or surpasses 1000 μ g/ml.The good bactericide of inhibition to trichoderma asperellum Thz01 mycelial growth has tpn and eugenol, its EC
50value is respectively 4.15 μ g/ml and 2.62 μ g/ml (table 1).
While being 1% by comparing inhibiting rate, the theoretical value of medicament is found with the drug concentration measured value that does not suppress mycelial growth completely: theoretical value and measured value are consistent substantially, if can filter out suitable mixed bactericide, measured value can be used as the reference value (table 2) of field test Chinese medicine application concentration.
The inhibition of table 1. medicament to trichoderma asperellum Thz01 mycelial growth
Table 2. does not suppress theoretical value and the measured value of trichoderma asperellum Thz01 mycelial growth
(2) inhibitory action of different bactericide bacterium to spore germination
The mould spore of wood that scraping is grown 8 days on PDA medium, prepares spore suspension, and being adjusted to concentration is 10
6individual/ml.Measure the concentration range that each Medicaments on Trichoderma viride spore germination starts inhibiting rate, then around this concentration 5 series concentration setting up and down.Reagent agent is made into respectively to certain density mother liquor, according to the concentration of having set, draws needed volume, join in the triangular flask that contains 45 ℃ of left and right PDA medium.After cooling, draw 100 μ l spore suspensions, drip in culture dish center, with triangular glass, be coated with rod coating evenly.At 25 ℃, cultivate after 12h, micro-Microscopic observation also calculates Germination suppression rate.Same, draw calculating virulence regression equation.Drug concentration while being 1% according to virulence regression equation calculating inhibiting rate arranges drug concentration gradient on the basis of this theoretical value, under identical training method and condition, determines and does not suppress the measured value of spore germination completely, and each concentration is processed and repeated 3 times.
Result shows, in 13 kinds of bactericide, Kocide SD and eugenol all do not have inhibition to spore germination; Three second aluminium phosphates and mandipropamid are poor to the inhibition of trichoderma asperellum Thz01; A Xi meter Da, dimethomorph mancozeb, second aluminium MnZn, tpn and the third gloomy iprovalicarb are obvious to the inhibition of trichoderma asperellum Thz01 spore germination, its EC
95concentration is all below 0.05 μ g/ml (table 3).
While being 1% by comparing inhibiting rate, the theoretical value of medicament is found with the drug concentration measured value that does not suppress spore germination completely: theoretical value and measured value are consistent substantially, if can filter out suitable mixed bactericide, measured value can be used as the reference value (table 4) of field trial Chinese medicine application concentration.
The inhibition of table 3. medicament to trichoderma asperellum Thz01 spore germination
Note: Kocide SD and eugenol do not have inhibitory action to spore germination; Amici reaches, dimethomorph mancozeb, second aluminium MnZn, tpn and the third gloomy iprovalicarb have strong inhibition effect to spore germination.Above 7 kinds of medicaments are not listed in table.
Table 4. does not suppress theoretical value and the measured value of trichoderma asperellum Thz01 spore germination
arepresent that bactericide has strong inhibitory action to spore germination, in the extremely low situation of concentration (c < 0.01), inhibiting rate is still more than 95%;
brepresent that bactericide does not have inhibitory action to spore germination, in dense situation (c > 1000), inhibiting rate is still below 5%.
(3) inhibitory action of different bactericide bacterium to Sporulation
Above two experiments have drawn respectively the measured value of every kind of medicament to the mycelia of trichoderma asperellum Thz01 and spore germination unrestraint concentration, by the comparison of these two values, select less value to make concentration and further detect the inhibitory action of bactericide to trichoderma asperellum Thz01 product spore.Get the Trichoderma mycelia piece of cultivating a day, mycelia surface is placed in sterile petri dish upward, adds bactericide liquid until surpass mycelia piece surface, contrast is to add equivalent sterile water to process, then be placed in 25 ℃ of incubators, place after 24h, examine under a microscope and produce spore quantity.
Result shows that the poor bactericide of trichoderma asperellum Thz01 product spore inhibition is had to mandipropamid, metalaxyl, flumorph and eugenol, and the mycelium surface sporulation quantity of processing through these bactericide is substantially identical with contrast; Trichoderma asperellum Thz01 produced to the good bactericide of spore inhibition have that Amici reaches, dimethomorph mancozeb, tpn, second aluminium MnZn, Kocide SD and three second aluminium phosphates, through the mycelium surface of these chemicals treatment, substantially do not produce spore (table 5).
The inhibition of table 5. medicament to trichoderma asperellum Thz01 Sporulation
+ representing to produce conidium on mycelium, plus sige more multilist shows that the conidium of generation is more
Above-mentioned 3 result of the tests show, mycelial growth, spore germination and product spore are different to the susceptibility of bactericide.The concentration that eugenol does not suppress trichoderma asperellum Thz01 mycelial growth is 1.5 μ g/ml, and the concentration that does not suppress to produce spore is 6 μ g/ml, and concentration is while reaching 200 μ g/ml, to spore germination also unrestraint.The concentration that mandipropamid does not suppress trichoderma asperellum Thz01 mycelial growth is 250 μ g/ml, and the concentration that does not suppress spore germination is 80 μ g/ml, and the concentration that does not suppress to produce spore is 160 μ g/ml.
Eugenol recommendation amount is aborning 30g/ mu, and being watered 60L(is that concentration is 1.5 μ g/ml), as can be seen here, directly use the concentration of using in producing also not suppress mycelial growth, spore germination and the product spore of trichoderma asperellum Thz01.Yet mandipropamid recommendation concentration is aborning 12g/ mu, is watered 30L(100 μ g/ml), under this concentration, the mycelium growth and sporulation unrestraint effect of mandipropamid to trichoderma asperellum Thz01, has to a certain extent and suppresses spore germination.
The wooden removing mildew of embodiment 3 and the mixed screening test in bactericide field
First divide community, field, arrange respectively blank district, medicament control treatment district and wood mould-medicament mixed processing district.During capsicum field planting, wood mould-medicament mixed processing district is transplanted to pepper seedling in planting hole jointly together with wooden removing mildew, 1.5-2 gram is used in every strain; Blank district and contrast medicament district do not use wooden removing mildew.After one month, with filling with root inoculation method inoculation phytophthora spore suspension, suspension concentration is 10
7individual/ml, every strain capsicum inoculation 3ml.Inoculate after 3 days, wood mould-medicament mixed processing district sprays bactericide according to following 6 amount of application gradients respectively: mandipropamid 6g/ mu, 6.6g/ mu, 7.2g/ mu, 7.8g/ mu, 8.4g/ mu, 9g/ mu, every mu is watered 30L; Eugenol 24g/ mu, 25g/ mu, 26g/ mu, 27g/ mu, 28g/ mu, 29g/ mu, every mu is watered 60L.Medicament control treatment district is according to application concentration mandipropamid 12g/ mu in producing, and every mu is watered 30L and eugenol 30g/ mu, and every mu is watered 60L and sprays bactericide.Each is processed and repeats 3 times, and each repeats 12 strains.Spray medicine is after 15 days, and strain incidence inoculated in record.
(1) after 15 days, respectively record incidence with spray medicine at the initial stage of a disease, and calculate the incidence of disease and disease index.
Stage division: 0 grade: asymptomatic; 1 grade: the slight blackening of seedling rhizome portion, blade is not wilted or restorability is wilted; 2 grades: the blackening of seedling rhizome portion reaches 1-2cm, blade irrecoverability is wilted, and lower blade is even to be had and come off; 3 grades: the blackening of seedling rhizome portion surpasses 2cm, and blade is obviously wilted or fallen leaves obviously; 4 grades: the seedling rhizome portion blackening contracting of hanging, all fallen leaves or plant wither except growing point; 5 grades: plant is withered; The incidence of disease and disease index calculate by formula (1), formula (2):
Diseased plant rate (%)=100 * diseased plant number/investigate total strain numerical expression (1)
Disease index=100 * ∑ (the sick numbers of sheets at different levels * typical values at different levels)/(investigating total number of sheets * highest typical value) formula (2)
(2) calculate drug effect
According to the disease index of each community, calculate medicament control treatment district and wood mould-control efficiency in medicament mixed processing district.Control efficiency is calculated by formula (3):
Control efficiency (%)=100 * (contrast disease index-processing disease index)/contrast disease index
Formula (3)
According to above result of the test, calculate wood mould-medicament mixes control efficiency and the control efficiency of bactericide use only.
Field biological and ecological methods to prevent plant disease, pests, and erosion result of the test shows, for trichoderma asperellum Thz01, when the mixing amount of application of 250g/L mandipropamid suspending agent (Switzerland Xian Zhengda crop protection Co., Ltd) is 7.8g/ mu, is watered 30L(active ingredient 1.95g; Active ingredient concentration 65 μ g/ml) time, control efficiency is 84.71%, is better than preventive effect 84.06%(Fig. 1 of the field of above-mentioned independent chemicals treatment).Under this consumption, mandipropamid reduces 4.2g/ mu than normal usage amount, and decrement bactericide is used 35%.In Fig. 1, figure A is dispenser disease incidence of disease measurement result after 15 days, and wooden removing mildew+7.8 gram/acre mandipropamid control efficiency is similar to independent chemical control efficiency; Figure B is dispenser disease index measurement result after 15 days, and wooden removing mildew+7.8 gram/acre mandipropamid control efficiency is similar to independent chemical control efficiency; Figure C is that dispenser prevents and treats statistical effect result after 15 days, and wooden removing mildew+7.8 gram/acre mandipropamid control efficiency is similar to independent chemical control efficiency.
When 0.3% soluble eugenol solvent (production of Baoding Ya Da Chemical Co., Ltd.) amount of application 27g/ mu, be watered 60L(active ingredient 0.081g; Active ingredient concentration 1.35 μ g/ml) time, control efficiency is 87.45%, is better than preventive effect 86.82%(Fig. 2 of the field of above-mentioned independent chemicals treatment).Under this consumption, eugenol reduces 3g/ mu than normal usage amount, and decrement bactericide is used 10%.In Fig. 2, figure A is dispenser disease incidence of disease measurement result after 15 days, and wooden removing mildew+27 gram/acre eugenol control efficiency is similar to independent chemical control efficiency; Figure B is dispenser disease disease index measurement result after 15 days, and wooden removing mildew+27 gram/acre eugenol control efficiency is similar to independent chemical control efficiency; Figure C is that dispenser prevents and treats statistical effect result after 15 days, and wooden removing mildew+27 gram/acre eugenol control efficiency is similar to independent chemical control efficiency.
Wood removing mildew and decrement bactericide mix under service condition, and because the usage amount of two kinds of bactericide is all lower than being used separately the amount of bactericide, therefore, under this disease control condition, wooden removing mildew is not subject to the impact of bactericide.
The inhibition test of embodiment 4 bactericide to phytophthora PC004 and trichoderma asperellum Thz01
Preparation contains the PDA dull and stereotyped (Φ=90 ㎜) that concentration is 65 μ g/ml mandipropamids and 1.35 μ g/ml eugenols respectively, take that to add the PDA flat board of equal-volume sterile water be contrast.With card punch (Φ=5 ㎜), from cultivating the Trichoderma bacterium colony edge of 2 days or cutting bacterium cake from the phytophthora blight of pepper bacterium colony edge of cultivating 5 days, be inoculated in containing medicament PDA medium central authorities, be placed in dark culturing in 25 ℃ of constant incubators.Each is processed and repeats 3 times.After trichoderma asperellum Thz013 days, measure colony diameter; After phytophthora blight of pepper PC0047 days, measure colony diameter.
Result shows, phytophthora blight of pepper PC004 cultivates after 7 days on the PDA flat board that contains the former medicine of 65 μ g/ml mandipropamid, grow completely suppressed, and the growth of trichoderma asperellum Thz01 is completely not suppressed, cultivates its bacterium colony size and contrast consistent (Fig. 3) after 3 days.Same, phytophthora blight of pepper PC004 cultivates after 7 days on the PDA flat board that contains the former medicine of 1.35 μ g/ml eugenol, grow completely suppressed, and the growth of trichoderma asperellum Thz01 is completely not suppressed, cultivate after 3 days its bacterium colony size with contrast consistent.
In Fig. 3, figure A, phytophthora blight of pepper PC004 cultivates after 7 days on the PDA flat board that contains the former medicine of 65 μ g/ml mandipropamid, grows completely suppressed; A1, A2, A3 are repetitions; CK is contrast.Figure B, trichoderma asperellum Thz01 cultivates after 3 days on the PDA flat board that contains the former medicine of 65 μ g/ml mandipropamid, and bacterium colony size is consistent with contrast; B1, B2, B3 are repetitions; CK is contrast.Figure C, phytophthora blight of pepper PC004 cultivates after 7 days on the PDA flat board that contains the former medicine of 1.35 μ g/ml eugenol, grows completely suppressed; C1, C2, C3 are repetitions; CK is contrast.Figure D, trichoderma asperellum Thz01 cultivates after 3 days on the PDA flat board that contains the former medicine of 1.35 μ g/ml eugenol, and bacterium colony size is consistent with contrast; D1, D2, D3 are repetitions; CK is contrast.
Claims (7)
1. a method for wooden removing mildew control mixed with bactericide capsicum epidemic disease in pepper planting process, the plantation of capsicum comprises sowing, grows seedlings, transplant planting, cultivate and gather, and it is characterized in that, described method comprises:
(1) while sowing, sowing after pepper seed is mixed with wooden removing mildew; Or, when Hot Pepper Seedling transplant planting, use wooden removing mildew;
(2) during pepper cultivation, before disease occurs or initial stage of origination use bactericide;
Wherein, the active bacterial strain in described wooden removing mildew is trichoderma asperellum (Trichoderma asperellum) Thz01, and the preserving number of trichoderma asperellum Thz01 is CGMCC No.6422;
Described bactericide is one or both in eugenol and mandipropamid; In every mu, the consumption of eugenol active ingredient is 0.078-0.084g; In every mu, the consumption of mandipropamid active ingredient is 1.8-2.1g.
2. method according to claim 1, is characterized in that, during pepper cultivation, within 0.5-1 month before disease occurs, at capsicum rhizosphere, uses wooden removing mildew around.
3. method according to claim 1 and 2, is characterized in that, the consumption of described wooden removing mildew is 1.5-2g/ strain.
4. method according to claim 1, it is characterized in that, described wooden removing mildew is prepared by the following method: trichoderma asperellum Thz01 spore suspension is inoculated in the wheat that boils sterilizing, in illumination in 12 hours and 12 hours dark alternate culture 15-20 days, makes wooden removing mildew.
5. method according to claim 4, is characterized in that, the viable bacteria concentration of described trichoderma asperellum Thz01 spore suspension is 10
7-10
8individual/mL.
6. method according to claim 4, is characterized in that, in every kilogram of wheat, the consumption of described trichoderma asperellum Thz01 spore suspension is 2-4mL.
7. method according to claim 4, is characterized in that, stirs in time wheat between culture period.
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| CN103392744A (en) * | 2013-06-24 | 2013-11-20 | 浙江大学 | Trichoderma preparation for controlling pepper phytophthora blight and field tank mix pesticide containing the same |
| CN104273125B (en) * | 2013-06-27 | 2016-03-02 | 浙江大学 | The application of 6-pentyl-2 hydrogen-pyran-2-one in control plant Oomycete disease |
| CN105432381B (en) * | 2015-12-07 | 2018-04-20 | 江苏省农业科学院 | A kind of preventing control method of capsicum epidemic disease |
| CN105638654A (en) * | 2015-12-23 | 2016-06-08 | 江西正邦生物化工有限责任公司 | Compound fungicide comprising Trichoderma asperellum spore powder of and application thereof |
| CN106472573B (en) * | 2016-10-12 | 2019-09-17 | 江西劲农作物保护有限公司 | A kind of trichoderma asperellum spore and cymoxanil combination sterilization composition |
| CN109287401A (en) * | 2018-11-28 | 2019-02-01 | 湖南省邵阳市拥思农副产品销售有限公司 | A kind of method of pepper planting |
| CN109868225B (en) * | 2019-04-16 | 2022-11-01 | 广西壮族自治区农业科学院植物保护研究所 | Trichoderma asperellum N-8-2 and application thereof |
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