CN111676143A - Application of Alternaria alternata (Alternaria alternata) in biocontrol of cucurbitaceae - Google Patents
Application of Alternaria alternata (Alternaria alternata) in biocontrol of cucurbitaceae Download PDFInfo
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Abstract
The invention discloses an application of Alternaria alternata (Alternaria alternata) in biocontrol of citrullinia seed, wherein the fermentation liquor of Alternaria alternata (Alternaria alternata) is adopted to have good inhibition effect on the germination determination of the citrullinia seed, and the inhibition rate of the fermentation liquor of Alternaria alternata (Alternaria alternata) and the crude extract of Alternaria alternata (Alternaria alternata) on the germination of the citrullinia seed can reach 100 percent by determining the inhibition effect of the fermentation liquor of Alternaria alternata (Alternaria alternata) and the crude extract of Alternaria alternata (Alternaria alternata) on the germination of the citrullinia seed and the test of inoculating the citrullinia seed with the fermentation liquor of Alternaria alternata (Alternaria alternata); the inhibition rate of the fermentation liquor of Alternaria alternata (Alternaria alternata) of 25 times of the dilution liquid on the seed of the; the wound of the plant is browned 2-3 days after the plant is inoculated with the Alternaria alternata (Alternaria alternata) fermentation liquor, and the whole plant is wilted and died 5-7 days later. The application of Alternaria alternata (Alternaria alternata) in the aspect of preventing and controlling the Guaranthus gourmet has obvious technical effect and application and popularization value.
Description
Technical Field
The invention relates to the technical field of microorganisms and application thereof, in particular to application of alternaria alternata (alternaria alternata) in biocontrol of cucurbitaceae.
Background
Orobanchhe aegyptiaca (Orobanchhe) is an annual herbaceous plant of the genus Orobanchaceae (Orobanchaceae). Due to the lack of leaves, chlorophyll and functional roots, broomrape is completely parasitic on the root system of a host plant, and maintains self growth by drawing water, nutrients and various growth hormones from the host, thereby causing serious harm to the host. Broomrape weeds have severely compromised agricultural production in many areas. The economic losses caused by the orobanche coerulescens worldwide are billions of dollars per year. In recent years, the brook outbreak in large area in Hebei, inner Mongolia, Xinjiang and other places seriously affects the yield and variety of crops, and causes huge economic loss to farmers. Among them, Xinjiang is the most serious area where the occurrence of Liaobang is most serious, and Liaobang is mainly parasitic on crops such as tomatoes, potatoes, eggplants, tobaccos and the like of the cucurbits and the solanaceae. According to the report, in Xinjiang in China, the area of the melon seriously damaged by Liantang is 5000-5500 hm2Processing tomato at 7000hm2. The parasitic rate of the cucurbitacin to the host is generally 30-40%, the parasitic rate of the cucurbitacin to the host is up to 100% in seriously damaged farmlands, and the growth of crops and the yield and the quality of fruits are seriously influenced. The generation of the orobanche coerulescens can reduce the yield of the melons and the watermelons by 20-70 percent and reduce the yield of the processed tomatoes by 30-80 percent.
The broomrape has a very short growth period, only about 30 days are needed for completing one growth period, each broomrape can generate 5-10 ten thousand seeds, and more seeds can reach 45 ten thousand seeds, and the broomrape has a large number of seeds, is fine and easy to spread, and is difficult to remove when falling into the field. The broomrape seeds survive in soil, keep the germination capacity for 5-10 years, and survive for more than 30 years under proper conditions. The seed germination and parasitism of the broomrape are finished underground and are not easy to be noticed, and when the broomrape parasitizes crops are discovered, hosts are seriously damaged. The existing methods for controlling the broomrape comprise cultivation control, physical control, chemical control, biological control, resistant hosts and the like. Measures such as crop rotation, fertilization, deep ploughing, adjustment of sowing time, soil exposure and the like can also reduce the rank hazards to a certain extent, but the measures are labor-consuming and time-consuming and have poor effects. In addition to transgenic crops that are resistant to chemical herbicides, existing chemical herbicides cannot distinguish between crops and orobanche. Soil fumigation is the most effective but costly and prohibited due to the risk to the environment. None of the above control methods can effectively eliminate the broomrape. Therefore, the effective control of the current herba Orobanches is still a worldwide problem.
Biological control is a control method that utilizes the interrelationship between species of organisms to inhibit one organism from another. Its advantages are less pollution, saving resources, high safety to broomrape host crops, less damage and high safety to human and animals. In recent years, studies on the control of orobanche coerulescens by microorganisms have been increasing, and many microorganisms having reported potential for controlling orobanche coerulescens, including Fusarium (Fusarium spp.), Rhizobium (Rhizobium spp.), arbuscular mycorrhizal fungi and Pseudomonas (Pseudomonas spp.) have been reported to have the ability to control orobanche coerulescens, and no report has been made on the use of Alternaria alternata (Alternaria alternata) in the control of orobanche coerulescens in the prior art. Compared with microorganisms introduced from other areas, the application of the local biocontrol bacteria resource is more suitable for the local ecological environment, and is beneficial to the colonization of the microorganisms and the continuous play of the control action.
Disclosure of Invention
Aiming at the technical problems that the existing strains have limited control range, low fermentation level, large control effect influenced by environment and the like in the control of the broomrape, no document and report is found to record the application of Alternaria alternata (Alternaria alternata) in the control of the broomrape. The invention aims to provide an application of Alternaria alternata (Alternaria alternata) in the biocontrol of the citrullinia seed, wherein the fermentation liquor of the Alternaria alternata (Alternaria alternata) has good inhibition effect on the determination of the germination of the citrullinia seed, and the germination inhibition rate of the original liquor of the fermentation liquor of the Alternaria alternata (Alternaria alternata) and the crude extract of the Alternaria alternata (Alternaria alternata) to the citrullinia seed can reach 100 percent by performing the activity determination on the citrullinia seed and the inoculation of the citrullinia seed plant test on the fermentation liquor of the Alternaria alternata (Alternaria alternata) with the crude extract of the Alternaria alternata (Alternaria alternata) at the concentration of 10 mu g/mL; the inhibition rate of the fermentation liquor of alternaria alternata (Alternariaalternata) of 25 times of the dilution liquid on the seed of; the wound of the plant is browned 2-3 days after the plant is inoculated with the Alternaria alternata (Alternaria alternata) fermentation liquor, and the whole plant is wilted and died 5-7 days later. Shows that the application of Alternaria alternata (Alternaria alternata) in the aspect of preventing and controlling the cucurbitaceous has obvious technical effect.
The invention provides application of Alternaria alternata (Alternaria alternata) in biocontrol of cucurbitaceae.
Further, the invention provides a culture method of Alternaria alternata (Alternaria alternata) applied to the biocontrol of cucurbitaceae, and the specific technical scheme is as follows:
(1) cutting small blocks of about 0.3cm at the boundary of the disease keys of a naturally-occurring Guaranga aethiopica plant by adopting purchased or self-screened Alternaria alternata (Alternaria alternata), adding 70% alcohol to soak for 1min, then soaking in a 3% sodium hypochlorite solution to disinfect for 3min, continuously rinsing with sterile water for 4 times, taking out the small blocks of the Guaranga aethiopica plant by using sterile tweezers, absorbing water on sterile filter paper, and then placing on a PDA culture medium flat plate to culture for 2-3 days in a 28 ℃ culture box; hyphae from the colony edge were inoculated into a new separation plate for purification, and purification was continued 4 times.
(2) Transferring purified and preserved Alternaria alternata (Alternaria alternata) into PDA plate, activating at 28 deg.C for 48 hr, transferring the activated test strain cake into liquid culture medium, and culturing at 28 deg.C for 135r min-1Performing shake culture for 24h to obtain seed liquid; inoculating 5mL of seed liquid into PDA liquid culture medium, and culturing at 28 deg.C for 135r min-1Performing shaking culture for 120h for later use; a piece of whatman filter paper (GA/A) is wetted by sterile water and placed in culture dishes, then 20 seeds of the cucurbitacin are placed on the filter paper, 2mLGR24 and 2mL of fermentation liquor are added into each culture dish, 2mL of GR24(GR24 is an artificial synthetic analogue of strigolactone) and 2mL of sterile water are added into a blank control, and 2mLGR24 and 2mLPDA liquid culture medium are added into PDCK (PDCK is a liquid culture medium as a control) treatment, and 3 times of repetition are carried out; culturing at 25 deg.C in dark for 7 days, and observing the seed under microscopeAnd (4) recording the germination condition and the germination quantity, calculating the correction inhibition rate, and screening strains with the correction inhibition rate of 100%.
Also, the present invention provides for the preparation of a semisolid seed liquid of Alternaria alternata (Alternaria alternata): transferring purified and preserved Alternaria alternata (Alternaria alternata) to PDA plate for 5d growth, adding the fungus cake into semisolid culture medium at 28 deg.C and 135r min-1Shaking and culturing for 24h to obtain semi-solid seed liquid.
In the present invention, it is preferable to add a cake of Alternaria alternata (Alternaria alternata) in a semi-solid medium at 28 ℃ and 135 r.min-1Shaking and culturing for 24h to obtain semi-solid seed liquid, wherein the semi-solid culture medium comprises 10g of peptone, 40g of maltose, 10g of yeast extract powder, 4g of agar and 1000mL of distilled water.
The present invention provides for the preparation of Alternaria alternata (Alternaria alternata) fermentation broths: transferring 5mL of seed liquid into rice culture medium with a pipette, and culturing at 28 deg.C for 15d to obtain Alternaria alternata (Alternaria alternata) fermentation liquid.
In the invention, the Alternaria alternata (Alternaria alternata) seed liquid is preferentially transferred into a rice culture medium, wherein the rice culture medium comprises 80g of rice, 1.2g of yeast powder and 120mL of distilled water, and the rice culture medium is sealed, soaked overnight and sterilized for 30min at 121 ℃.
The present invention provides for the preparation of crude extracts of Alternaria alternata (Alternaria alternata): mechanically pulverizing rice and solid fermentation liquid, adding 240mL ethyl acetate into each bottle, soaking, ultrasonically extracting for 2 times, and rotary evaporating to obtain Alternaria alternata (Alternaria alternata) crude extract.
Further, the invention provides application of Alternaria alternata (Alternaria alternata) fermentation liquor in preventing and controlling the citrullinia cupreum.
The invention provides application of a crude extract of Alternaria alternata (Alternaria alternata) in controlling citrullus lanatus.
The invention provides application of Alternaria alternata (Alternaria alternata) fermentation liquor in different inoculation modes of melons in preventing and treating melon broomrape.
In the invention, Alternaria alternata (Alternaria alternata) is a harmless and safe strain allowed to be used by the department of agriculture, is purchased from China general microbiological culture Collection center (CGMCC) and can be obtained by common technicians from public sources, the culture of the Alternaria alternata (Alternaria alternata) adopts a known culture medium and culture method, and PDA solid, liquid culture medium and PCA culture medium are adopted without limiting the implementation of the invention.
The Alternaria alternata (Alternaria alternata) separated by the technical scheme is applied to the aspect of preventing and controlling cucurbiturils, and has the following beneficial effects:
the invention provides application of Alternaria alternata (Alternaria alternata) in biocontrol of cucurbitaceae. The test of inhibiting the germination of the seeds of the Orobanchum by adopting the fermentation liquor of the Alternaria alternata has good inhibiting effect, and the germination inhibiting rate of the seeds of the Orobanchum reaches 100 percent by determining the inhibiting effect of the fermentation liquor of the Alternaria alternata and the crude extract of the Alternaria alternata on the germination of the seeds of the Orobanchum and inoculating the Orobanchum plants by the fermentation liquor of the Alternaria alternata, wherein the original liquor of the fermentation liquor of the Alternaria alternata and the crude extract with the concentration of 10 mu g/mL; the inhibition rate of the fermentation liquor of Alternaria alternata (Alternaria alternata) of 25 times of the dilution liquid on the seed of the; the wound of the plant is browned after the plant is inoculated with the alternaria alternata (Alternaalternala) fermentation liquor for 2-3 days, and the whole plant is wilted and died after 5-7 days. Shows that the application of Alternaria alternata (Alternaria alternata) in the aspect of preventing and controlling the cucurbitaceous has obvious technical effect.
Drawings
FIG. 1 is a graph showing the effect of a fermentation broth of Alternaria alternata (Alternaria alternata) on the inhibition of germination of seeds of Orobanches, in which A: comparison; b: 25 times of fermentation liquor; c: 50 times of fermentation liquor.
FIG. 2 is a graph showing the effect of crude extracts of Alternaria alternata (Alternaria alternata) on the inhibition of germination of seeds of Orobanches, in which A: comparison; b: and (4) crude extraction.
FIG. 3 is a graph showing the effect of different inoculation patterns of Alternaria alternata (Alternaria alternata) fermentation broth on the pathogenesis of Guaran, wherein A: carrying out acupuncture inoculation; b: spraying and inoculating; c: smearing and inoculating; d: blank control.
Detailed Description
The present invention will be described by way of examples, but the present invention is not limited to the examples.
All reagents and equipment selected for use in the present invention are well known in the art and are not intended to limit the practice of the present invention, and other reagents and equipment well known in the art may be suitable for use in the practice of the following embodiments of the present invention.
The materials of the invention are: alternaria alternata (Alternaria alternata) is available from public sources, and equipment and instruments used in the process are all equipment common in the art.
The culture of Alternaria alternata (Alternaria alternata) according to the invention is preferably prepared using a culture medium according to the following formulation:
PDA culture medium: 200g of potato, 17g of glucose, 20g of agar and 1000mL of distilled water.
PCA culture medium: 2.5g of yeast powder, 5g of peptone, 1g of glucose, 15g of agar and 1000mL of distilled water, and the pH value is 7.0.
Example 1: application of Alternaria alternata (Alternaria alternata) fermentation liquor in preventing and treating cucurbitaceous papyriferus
The application of the Alternaria alternata (Alternaria alternata) fermentation liquid in preventing and controlling the citrullus lanatus comprises the following specific steps: growing exogenously or self-screened Alternaria alternata strain on PDA plate for 5 days, adding the bacterial cake into semisolid culture medium (formula: 10g of peptone, 40g of maltose, 10g of yeast extract powder, 4g of agar and 1000mL of distilled water), and performing temperature control at 28 ℃ and 135 r.min-1And shaking and culturing for 24h to obtain the semi-solid seed liquid. Adding 5mL of semi-solid seed solution into rice culture medium (formula: rice 80g, yeast powder 1.2g and distilled water 120 mL), sealing, soaking overnight, sterilizing at 121 deg.C for 30min, and standing at 28 deg.C for 15 d.
Respectively diluting the fermentation liquor of Alternaria alternata (Alternaria alternata) to 25 times and 50 times, testing the inhibition effect of the fermentation liquor with different concentrations on the germination of the seeds of the orobanum cucurbitacearum, and obtaining the data in table 1 that the inhibition rate of the stock solution of the fermentation liquor of the Alternaria alternata (Alternaria alternata) on the germination of the seeds of the orobanum cucurbitacearum is 100%; when the fermentation broth of alternaria alternata (alternaria alternata) is diluted 25-fold, the corrected inhibition rate of the fermentation broth to the germination of the seeds of the citrulline is measured to be 91.34%; when the fermentation liquor of Alternaria alternata (Alternaria alternata) is diluted by 50 times, the corrected inhibition rate of the fermentation liquor of Alternaria alternata (Alternaria alternata) on the germination of the seeds of the citrullinia cupra is measured to be 87.68%, and therefore the dilution liquor of the fermentation liquor of Alternaria alternata (Alternaria alternata) with different times has a good inhibition effect on the germination of the seeds of the citrullinia cupra. Meanwhile, the germination effect of the Alternaria alternata (Alternaria alternata) fermentation liquid on the orobanche sonchifolia seeds can be observed through microscope observation, as shown in the attached drawing 1, it can be seen from the attached drawing 1 that the orobanche sonchifolia seeds treated by the Alternaria alternata (Alternaria alternata) fermentation liquid do not germinate, but the growth of the germ tubes of the orobanche sonchifolia seeds in a control group is good.
Table 1: germination effect of Alternaria alternata (Alternaria alternata) fermentation broth on seeds of orobanum cucurbitacearum
Note: PDCK is liquid medium as control and CK is sterile water control.
Example 2: application of crude extract of Alternaria alternata (Alternaria alternata) in preventing and treating cucurbitaceous peperomia
Fermenting the strain: growing exogenously or self-screened Alternaria alternata strain on PDA plate for 5 days, adding the bacterial cake into semisolid culture medium (formula: 10g of peptone, 40g of maltose, 10g of yeast extract powder, 4g of agar and 1000mL of distilled water), and performing temperature control at 28 ℃ and 135 r.min-1And shaking and culturing for 24h to obtain the semi-solid seed liquid. Adding 5mL of semi-solid seed solution into rice culture medium (formula: rice 80g, yeast powder 1.2g and distilled water 120 mL), sealing, soaking overnight, sterilizing at 121 deg.C for 30min, and standing at 28 deg.C for 15 d.
Crude extract of the strain: mechanically pulverizing rice and thallus, soaking in 240mL ethyl acetate per bottle, ultrasonic extracting for 2 times, and rotary evaporating to obtain crude extract.
And (3) determining the germination inhibition effect of the crude extract of the strain on the seed of the orobanche cucurbitacearum: 20 sterile dried seeds of Orobanches, 0.5mL of strigolactone (GR24), and 0.5mL of sterile water were added to each well of a 24-well plate, 10. mu.g of the crude extract was added to the treatment group, and sterile water was added to the control group, and the treatment was repeated 3 times. And (4) after the seeds are cultured for 5 days at 25 ℃ in a shading mode, observing the germination quantity of the seeds of the orobanche coerulescens under a body type microscope and recording the germination quantity.
Table 2: germination effect of Alternaria alternata (Alternaria alternata) crude extract on seed of Orobanchum schoenbergii
| Test strains | Percentage of germination (%) | Inhibition ratio (%) |
| 10 mu g/mL crude extract | 0 | 100 |
| CK | 90 | --- |
Note: CK is sterile water control.
As can be seen from the results in Table 2, the inhibition rate of germination of seeds of Orobanches was 100% when the crude extract of Alternaria alternata (Alternaria alternata) was 10. mu.g/mL. As shown in the attached figure 2, it is obvious that the seed of the orobanche japonica after the treatment of the crude extract of Alternaria alternata (Alternaria alternata) does not germinate under a microscope, while the germ tube of the seed of the orobanche japonica of the control group grows well.
Example 3: application of Alternaria alternata (Alternaria alternata) fermentation liquor in different inoculation modes in control of Orobanchum cucurbitacearum
The application of the Alternaria alternata (Alternaria alternata) fermentation liquor in different inoculation modes for preventing and treating the citrulline includes the following specific steps: inoculating on the plant of the orobanche cucurbitacearum by three modes of needling, smearing and spraying, wherein a needling method comprises the following steps: dipping the spore liquid with an inoculating needle, and needling the base part, the middle part and the top part of the broomrape plant respectively; the smearing method comprises the following steps: dipping the spore liquid by using a coater, and respectively coating three parts of the broomrape plant; after inoculation, the cotton is covered with wet absorbent cotton, and the cotton is covered with a sealing bag sprayed with sterile water for moisture preservation for 24h, and then the sealing bag and the absorbent cotton are removed. Spraying method: spraying spore liquid on the broomrape which just comes out of the soil, covering the broomrape with a sealing bag, preserving moisture for 24 hours, and then picking off the broomrape; blank control was sterile water, all 3 replicates. The study of the case of the broomrape disease started after 2d, and ended after 7d, the results of the study are shown in FIG. 3. As shown in the attached figure 3, the three methods can cause diseases of the Liequus cucurbitacearum plant, the wound turns brown after 2 to 3 days, the whole plant dies due to wilting after 5 to 7 days, and the contrast plant grows well.
The above examples show that, in the invention, a strain of Alternaria alternata (Alternaria alternata) is separated and screened from stems of naturally-occurring citrulline in a tomato field, and both the fermentation liquor and the crude extract of the strain have a significant inhibition effect on germination of citrulline seeds and have strong pathogenicity on citrulline plants. The germination of the citrulline seed is measured by Alternaria alternata (Alternaria alternata) fermentation liquor and crude extract, the germination inhibition rate of the raw liquor of the Alternaria alternata (Alternaria alternata) fermentation liquor and the crude extract with the concentration of 10 mu g/mL to the citrulline seed is 100%, the inhibition rate of the fermentation liquor of the Alternaria alternata (Alternaria alternata) is 91.34% when the fermentation liquor is diluted by 25 times, the inhibition rate of the fermentation liquor of the Alternaria alternata seed is 87.68% when the fermentation liquor of the Alternaria alternata (Alternaria alternata) is diluted by 50 times, and the bacterial strain is used for efficiently inhibiting the germination of the citrulline seed to prepare the pesticide bacteriostatic agent, so that the bacterial strain has wide application value in the technical field of microbial strain application.
The above examples are merely illustrative for clearly illustrating the present invention and are not intended to limit the embodiments. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. And obvious variations or modifications can be made while remaining within the scope of the present invention.
Claims (9)
1. Use of Alternaria alternata (Alternaria alternata) in biocontrol of citrullinia.
2. A culture method of Alternaria alternata (Alternaria alternata) applied to the biocontrol of Guaranga aequalata is characterized by comprising the following specific technical scheme:
(1) cutting small blocks of about 0.3cm at the boundary of the disease keys of a naturally-occurring Guaranga aethiopica plant by adopting purchased or self-screened Alternaria alternata (Alternaria alternata), adding 70% alcohol to soak for 1min, then soaking in a 3% sodium hypochlorite solution to disinfect for 3min, continuously rinsing with sterile water for 4 times, taking out the small blocks of the Guaranga aethiopica plant by using sterile tweezers, absorbing water on sterile filter paper, and then placing on a PDA culture medium flat plate to culture for 2-3 days in a 28 ℃ culture box; picking hyphae from the edge of the colony, inoculating the hyphae into a new separation plate for purification, and continuously purifying for 4 times;
(2) transferring purified and preserved Alternaria alternata (Alternaria alternata) into PDA plate, activating at 28 deg.C for 48 hr, transferring the activated test strain cake into liquid culture medium, and culturing at 28 deg.C for 135r min-1Performing shake culture for 24h to obtain seed liquid; inoculating 5mL of seed liquid into PDA liquid culture medium, and culturing at 28 deg.C for 135r min-1Performing shaking culture for 120h for later use; wetting a piece of whatman filter paper (GA/A) with sterile water, placing the filter paper in culture dishes, then placing 20 seeds of the cucurbitacin in the filter paper, adding 2mLGR24 and 2mL of fermentation liquor in each culture dish, adding 2mL of GR24 and 2mL of sterile water in a blank control, adding 2mLGR24 and 2mL of PDA liquid culture medium in PDCK treatment, and repeating for 3 times; culturing for 7d at 25 ℃ in the dark, observing the germination condition of the seed of the orobanche coerulescens in a body type microscope, recording the germination quantity, calculating the correction inhibition rate, and screening strains with the correction inhibition rate reaching 100%.
3. Preparation of solid seed liquid from Alternaria alternata (Alternaria alternata) for use in biocontrol of melon and fruit trees according to claim 1 or 2, characterized in that after transfer of purified and preserved Alternaria alternata (Alternaria alternata) onto PDA plates for 5d growth, the cake is taken and added to a semi-solid medium at 28 ℃, 135 r-min-1Shaking and culturing for 24h to obtain semi-solid seed liquid.
4. The method of claim 3, wherein the semi-solid seed liquid is prepared by adding a cake of Alternaria alternata (Alternaria alternata) to a semi-solid medium at 28 ℃ and 135 r-min-1Shaking and culturing for 24h to obtain semi-solid seed liquid, wherein the semi-solid culture medium comprises 10g of peptone, 40g of maltose, 10g of yeast extract powder, 4g of agar and 1000mL of distilled water.
5. The preparation of fermentation broth for Alternaria alternata (Alternaria alternata) in biocontrol of citrulline according to any of claims 1 or 2, characterized in that 5mL of seed solution is taken by a pipette and transferred into rice culture medium, and cultured for 15d at 28 ℃ to obtain Alternaria alternata (Alternaria alternata) fermentation broth.
6. The preparation method of fermentation liquor of Alternaria alternata (Alternaria alternata) in biocontrol of citrulline as claimed in claim 5, characterized in that Alternaria alternata (Alternaria alternata) seed liquid is transferred into rice culture medium, wherein the rice culture medium comprises 80g of rice, 1.2g of yeast powder and 120mL of distilled water, and the rice culture medium is sterilized at 121 ℃ for 30min after being sealed and soaked overnight.
7. Preparation of crude extract of Alternaria alternata (Alternaria alternata) for use in biocontrol of melon orobanche according to any of claims 1 or 2, characterized in that rice and solid fermentation broth are mechanically pulverized, soaked in 240mL of ethyl acetate per bottle, extracted with ultrasound 2 times, and rotary evaporated to obtain an extract of Alternaria alternata (Alternaria alternata) crude extract.
8. Use of a fermentation broth of Alternaria alternata (Alternaria alternata) according to claim 5 for controlling citrulline.
9. Use of a crude Alternaria alternata (Alternaria alternata) extract according to claim 7 for controlling Orobanchum cucurbitacearum.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114009452A (en) * | 2021-11-01 | 2022-02-08 | 新疆农业科学院植物保护研究所 | Application of fermentation broth of alternaria alternata JTF001 in inhibition of germination of seeds of orobanum cucurbitacearum |
| CN114806896A (en) * | 2022-06-06 | 2022-07-29 | 青岛农业大学 | Alternaria alternata, herbicide and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102191184A (en) * | 2011-04-08 | 2011-09-21 | 中国计量学院 | Biocontrol endophytic fungi-Alternaria alternata |
| CN102204570A (en) * | 2011-04-08 | 2011-10-05 | 中国计量学院 | Application of alternaria alternate metabolic products in cucumber rhizoctonia solani prevention and control |
| CN102217656A (en) * | 2011-04-08 | 2011-10-19 | 中国计量学院 | Application of alternaria alternata metabolite to controlling Botrytis cirerea |
| CN109897788A (en) * | 2019-03-08 | 2019-06-18 | 浙江工业大学 | A kind of rape rod method and preparing the application in bacteriostatic agent |
-
2020
- 2020-06-29 CN CN202010608274.4A patent/CN111676143A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102191184A (en) * | 2011-04-08 | 2011-09-21 | 中国计量学院 | Biocontrol endophytic fungi-Alternaria alternata |
| CN102204570A (en) * | 2011-04-08 | 2011-10-05 | 中国计量学院 | Application of alternaria alternate metabolic products in cucumber rhizoctonia solani prevention and control |
| CN102217656A (en) * | 2011-04-08 | 2011-10-19 | 中国计量学院 | Application of alternaria alternata metabolite to controlling Botrytis cirerea |
| CN109897788A (en) * | 2019-03-08 | 2019-06-18 | 浙江工业大学 | A kind of rape rod method and preparing the application in bacteriostatic agent |
Non-Patent Citations (3)
| Title |
|---|
| JOEL D M ET AL: "Biology and Management of Weedy Root Parasites[M].", 《HORTICULTURAL REVIEWS-WESTPORT THEN NEW YORK》 * |
| 姚兆群等: "新疆列当的种类、分布及其防治技术研究进展", 《生物安全学报》 * |
| 陈杰等: "利用微生物防除根寄生杂草列当", 《中国生态农业学报》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114009452A (en) * | 2021-11-01 | 2022-02-08 | 新疆农业科学院植物保护研究所 | Application of fermentation broth of alternaria alternata JTF001 in inhibition of germination of seeds of orobanum cucurbitacearum |
| CN114806896A (en) * | 2022-06-06 | 2022-07-29 | 青岛农业大学 | Alternaria alternata, herbicide and application thereof |
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