CN109845762B - Biological control agent combining trichoderma harzianum and azadirachtin and application thereof in prevention and control of soybean epidemic disease - Google Patents
Biological control agent combining trichoderma harzianum and azadirachtin and application thereof in prevention and control of soybean epidemic disease Download PDFInfo
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- 235000010469 Glycine max Nutrition 0.000 title claims abstract description 37
- 241000223260 Trichoderma harzianum Species 0.000 title claims abstract description 36
- 239000005878 Azadirachtin Substances 0.000 title claims abstract description 31
- VEHPJKVTJQSSKL-UHFFFAOYSA-N azadirachtin Natural products O1C2(C)C(C3(C=COC3O3)O)CC3C21C1(C)C(O)C(OCC2(OC(C)=O)C(CC3OC(=O)C(C)=CC)OC(C)=O)C2C32COC(C(=O)OC)(O)C12 VEHPJKVTJQSSKL-UHFFFAOYSA-N 0.000 title claims abstract description 31
- FTNJWQUOZFUQQJ-IRYYUVNJSA-N azadirachtin A Natural products C([C@@H]([C@]1(C=CO[C@H]1O1)O)[C@]2(C)O3)[C@H]1[C@]23[C@]1(C)[C@H](O)[C@H](OC[C@@]2([C@@H](C[C@@H]3OC(=O)C(\C)=C/C)OC(C)=O)C(=O)OC)[C@@H]2[C@]32CO[C@@](C(=O)OC)(O)[C@@H]12 FTNJWQUOZFUQQJ-IRYYUVNJSA-N 0.000 title claims abstract description 31
- FTNJWQUOZFUQQJ-NDAWSKJSSA-N azadirachtin A Chemical compound C([C@@H]([C@]1(C=CO[C@H]1O1)O)[C@]2(C)O3)[C@H]1[C@]23[C@]1(C)[C@H](O)[C@H](OC[C@@]2([C@@H](C[C@@H]3OC(=O)C(\C)=C\C)OC(C)=O)C(=O)OC)[C@@H]2[C@]32CO[C@@](C(=O)OC)(O)[C@@H]12 FTNJWQUOZFUQQJ-NDAWSKJSSA-N 0.000 title claims abstract description 30
- 241001646398 Pseudomonas chlororaphis Species 0.000 title claims abstract description 18
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- 241000233614 Phytophthora Species 0.000 claims abstract description 11
- NHOIBRJOQAYBJT-IMGVWCFESA-N nimbin Chemical compound C=1([C@@H]2C[C@H]3O[C@H]4[C@](C3=C2C)(C)[C@@H]([C@]2(C(=O)C=C[C@](C)([C@@H]2[C@H]4OC(C)=O)C(=O)OC)C)CC(=O)OC)C=COC=1 NHOIBRJOQAYBJT-IMGVWCFESA-N 0.000 claims abstract description 9
- ZQIYJHBQRBBBRZ-UHFFFAOYSA-N nimbin Natural products COC(=O)CC1C2C(C(OC(=O)C)C3OC4CC(C(=C4C13C)C)c5cocc5)C(C)(C=CC2=O)C(=O)OC ZQIYJHBQRBBBRZ-UHFFFAOYSA-N 0.000 claims abstract description 9
- 244000052616 bacterial pathogen Species 0.000 claims description 7
- 239000012681 biocontrol agent Substances 0.000 claims description 5
- 239000002552 dosage form Substances 0.000 claims description 5
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- XSTXAVWGXDQKEL-UHFFFAOYSA-N Trichloroethylene Chemical compound ClC=C(Cl)Cl XSTXAVWGXDQKEL-UHFFFAOYSA-N 0.000 description 1
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- 125000000917 azadirachtin group Chemical group 0.000 description 1
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Abstract
The invention discloses a trichoderma harzianum and azadirachtin combined biological control agent and application thereof in soybean epidemic prevention and control, belonging to the technical field of plant protection; the dosage of the nimbin and the trichoderma harzianum in the biological control agent is carried out according to the following conditions: the concentration of the azadirachtin is 0.70mg/L-0.90mg/L, and the concentration of the trichoderma harzianum is 1.0 multiplied by 106spore/mL; according to the invention, trichoderma harzianum and azadirachtin are compounded into the biological control agent for the first time and are applied to the control of soybean phytophthora blight, and data show that the biological control agent compounded by trichoderma harzianum and azadirachtin has a synergistic effect on the control of soybean phytophthora blight, the final concentration of the azadirachtin is 0.80mg/L, and the concentration of the trichoderma harzianum is 1.0 multiplied by 106The biological control agent has the most obvious inhibition effect on soybean epidemic disease when the spore is one/mL.
Description
Technical Field
The invention belongs to the technical field of plant protection, and particularly relates to a trichoderma harzianum and azadirachtin combined biological control agent and application thereof in prevention and control of soybean epidemic diseases.
Background
The pathogenic bacteria of soybean Phytophthora sojae is Phytophthora sojae (Phytophthora sojae). The disease is a soil-borne disease with extremely serious harm and is an important foreign quarantine object in China. The soybean phytophthora blight was discovered in indiana in the united states at the earliest time in 1948, and gradually occurs in the united states and in many countries in the world, phytophthora sojae is first isolated from shenchoo in northeast soybean production areas in China until 1989, and the disease also occurs in part of soybean production areas in China and causes serious harm at present. The phytophthora sojae can survive in soil for a long time to cause rotting of soybean seeds, roots and stems, and can be infected in the whole growth period of soybeans, the yield is generally reduced by 10-30%, the yield is seriously reduced by 50-70%, and some soybean diseases are even completely harvested. If the soybeans are infected before sprouting, seeds can be rotten, and cotyledons do not open and die finally after sprouting; if the soybean is infected in the adult plant period, water stain-like brown spots appear at the stem nodes initially, the diseased spots expand upwards and downwards, leaves on the upper part and the lower part of the plant gradually turn yellow and wither, the diseased part shrinks, the diseased spots gradually turn black brown, the roots appear water stain-like, the root system is dysplastic, and the plant is withered and dead; the symptoms of the soybean infection in the later growth stage are mainly manifested in that the lower leaves are yellow and dry, but are not easy to fall off, the scabs near the base and stem of the ground are dark brown, the stem is dry and easy to break, and the pods and the grains are easy to shrink. The phytophthora sojae has a complex and multi-cycle life history, and once the phytophthora sojae is introduced, the phytophthora sojae can survive in a disease area for a long time and is difficult to cure radically.
The nimbin has the characteristics that: neem (Azadirachta indica a. juss) is an arbor of the meliaceae family, belonging to the meliaceae genus, and is widely planted in tropical and subtropical areas; native to Burma and India, distributed and planted in more than 70 countries, and the highest yield in Asian countries such as India; neem is praised by the United states department of agriculture as a "tree that can solve global problems". To date, over 100 compounds have been discovered from neem seeds, bark, and fruits, at least 70 of which are biologically active, diterpenes, triterpenes, pentatriterpenes, and non-terpenoids. Mainly contains azadirachtin, azadirachtin triol, azadirachtin and the like, wherein the most important active substance is azadirachtin. The azadirachtin has the effects of inhibiting pests and mites and also inhibiting plant pathogenic bacteria, and is used for preventing and treating various diseases such as leaf spot, downy mildew, powdery mildew and the like of fruit trees and crops.
The existing soybean epidemic prevention and control agent is limited by the safety of crops, and the single agent of the soybean epidemic prevention and control agent cannot completely and effectively control the occurrence of soybean epidemic; the compound of the soybean epidemic disease preventing and treating agent can solve the problems of large toxicity and poor effect of a single pesticide, has less investment and short research and development period, can delay the generation of drug resistance of an agent by using different pesticides interactively, and has ideal use effect, so the compound of the pesticides with different action mechanisms is a research hotspot in the field of pesticides at present. In the prior art, no report related to the compounding of trichoderma harzianum and azadirachtin is provided.
Disclosure of Invention
1. Problems to be solved
Aiming at the problem that the existing soybean epidemic disease control agent is not compounded with azadirachtin into a biological control agent, the invention provides a biological control agent combining trichoderma harzianum and azadirachtin and application thereof in soybean epidemic disease control, and the biological control agent can effectively inhibit soybean phytophthora.
2. Technical scheme
In order to solve the above problems, the present invention adopts the following technical solutions.
A biocontrol agent combining Trichoderma harzianum and azadirachtin, wherein Trichoderma harzianum (Trichoderma harzianum) is purchased from Guangdong province collection of microorganisms under accession number GIM 3.442; the dosages of the nimbin and the trichoderma harzianum are carried out according to the following conditions: the concentration of the azadirachtin is 0.70mg/L-0.90mg/L, and the concentration of the trichoderma harzianum is 1.0 multiplied by 106spores/mL.
Preferably, the dosages of the nimbin and the trichoderma harzianum are carried out according to the following conditions: the concentration of the azadirachtin is 0.80 mg/L.
An application of the biological control agent combining trichoderma harzianum and azadirachtin in controlling soybean epidemic disease.
Preferably, the dosage form of the biocontrol agent is a pharmaceutically acceptable dosage form.
Preferably, the pathogenic bacteria of soybean Phytophthora blight is Phytophthora sojae (Phytophthora sojae).
3. Advantageous effects
Compared with the prior art, the invention has the beneficial effects that:
according to the invention, trichoderma harzianum and azadirachtin are compounded into a biological control agent for the first time and are applied to the control of soybean epidemic disease; the data show that the biological control agent compounded by trichoderma harzianum and azadirachtin has a synergistic effect on the control of soybean phytophthora blight, the final concentration of the azadirachtin is 0.80mg/L, and the concentration of the trichoderma harzianum is 1.0 multiplied by 106The biological control agent has the most obvious inhibition effect on soybean epidemic disease when the spore is one/mL.
Detailed Description
Example 1
Test materials:
(1) biocontrol fungus
The Trichoderma harzianum (Trichoderma harzianum) is purchased from Guangdong province culture Collection, and the preservation number is GIM 3.442.
(2) Chemical agent
Nimbin, dosage form: aqueous, manufacturer: shaanxi Sinot Biotechnology Ltd.
(3) Target pathogenic bacteria
The Phytophthora sojae (Phytophthora sojae) is provided by plant pathology research laboratory of university of agriculture in Anhui, and the host is soybean.
The experimental method comprises the following steps:
the operation is carried out by referring to a "synergistic coefficient method" disclosed in an issued patent (application No. 201310148580.4, granted publication No. CN103255198B, inventive subject matter: method for determining the synergistic effect of biocontrol fungi and chemical agents on target pathogenic bacteria, applicant: plant protection research institute of agriculture and forestry academy of sciences of Hebei province):
(1) inoculating the biocontrol fungus and the target pathogenic bacteria into a culture medium containing chemical agents at the same time with the same inoculation amount for opposite culture, measuring the colony radius after culturing for a period of time, and respectively using R as the colony radius of the biocontrol fungus and the target pathogenic bacteriaTtAnd RBtRepresents;simultaneously setting contrast culture, respectively inoculating the biocontrol fungi and the target pathogenic bacteria into the same culture medium which does not contain the chemical agent and is used for the confrontation culture in the same inoculation amount as the confrontation culture, culturing under the same culture condition and culture time as the confrontation culture, measuring the colony radius after the culture is finished, and respectively using R as the colony radius of the biocontrol fungi and the target pathogenic bacteriaTCKAnd RBCKRepresents;
(2) calculating the synergy coefficient according to formula (I):
co-operative coefficient (S) ═ RBt–RBCK)/(RTt–RTCK) Formula (I)
If S is more than or equal to 1.5, the biocontrol fungus and the chemical agent have synergistic action, if S is more than or equal to 1 and less than 1.5, the biocontrol fungus and the chemical agent have additive action, and if S is less than 1, the biocontrol fungus and the chemical agent have antagonistic action.
The specific operation of this embodiment is as follows:
accurately measuring chemical agent (azadirachtin), sucking 2mL of liquid medicine with a pipette, adding into sterilized 18mL of PDA plate, and making into PDA plate with different final concentrations (0.60mg/L, 0.70mg/L, 0.80mg/L, 0.90mg/L, 1.80 mg/L) of chemical agent; the same amount of inoculum (1.0X 10 final concentration after inoculation) was applied to the PDA plates containing the chemical agent6spore/mL) is inoculated with Trichoderma harzianum subjected to activation culture (constant temperature of 25 ℃, PDA plate, 3d) and Phytophthora sojae subjected to activation culture (constant temperature of 25 ℃, PDA plate, 3d) at the same time, and the culture is carried out in opposition; meanwhile, contrast culture is set, and trichoderma harzianum and phytophthora sojae are inoculated to the PDA plate without chemical agent in the same inoculation amount as the contrast culture. Placing upside down in a constant temperature incubator at 25 ℃ for 3 d. Each concentration treatment was repeated 5 times and the experiment was repeated 3 times. Measuring the radius of each processed colony after finishing the culture, taking an average value, and calculating a synergistic coefficient S according to a formula (I);
wherein R isBtShowing the relative radius of soybean phytophthora in the confronting culture groups,
wherein R isTtShowing the relative radius of the Trichoderma harzianum colonies in the opposing culture group,
wherein R isBCKRepresents the phytophthora sojae colony radius in the control culture group,
wherein R isTCKRepresents the radius of the Trichoderma harzianum colony in the control culture group;
in the above formula, the subtraction of the control value is to eliminate the test error;
TABLE 1
Furthermore, RBCKIs 3.34cm, RTCKIs 3.48 cm;
according to the judgment result of the 'synergy coefficient method':
when the nimbin and the trichoderma harzianum are used together, the mass concentration of the nimbin is in the range of 0.70mg/L-0.90mg/L, the synergistic coefficient to the phytophthora sojae is 1.75-5.59, and the synergistic effect is achieved. Taking into account, selecting azadirachtin with final concentration of 0.80mg/L and final concentration of 1.0 × 106The spores/mL of trichoderma harzianum are combined, and the biological control agent has the most obvious inhibition effect on the phytophthora sojae at the moment.
While the invention has been described in further detail in connection with specific embodiments thereof, it will be understood that the invention is not limited thereto, and that various other modifications and substitutions may be made by those skilled in the art without departing from the spirit of the invention, which should be considered to be within the scope of the invention as defined by the appended claims.
Claims (4)
1. An application of a biological control agent combining trichoderma harzianum and azadirachtin in controlling soybean epidemic disease is characterized in that: wherein said Trichoderma harzianumTrichoderma harzianumPurchased from Guangdong province culture Collection of microorganisms under the number GIM 3.442; the dosages of the nimbin and the trichoderma harzianum are carried out according to the following conditions: the concentration of the azadirachtin is 0.70mg/L-0.90mg/L, and the concentration of the trichoderma harzianum is 1.0 multiplied by 106spores/mL.
2. The use of a combination biocontrol agent from trichoderma harzianum and azadirachtin as claimed in claim 1 for the control of soybean phytophthora blight, characterized in that: the dosages of the nimbin and the trichoderma harzianum are carried out according to the following conditions: the concentration of the azadirachtin is 0.80 mg/L.
3. The use of a combination biocontrol agent from trichoderma harzianum and azadirachtin as claimed in claim 1 for the control of soybean phytophthora blight, characterized in that: the dosage form of the biological control agent is a pharmaceutically acceptable dosage form.
4. The use of a combination biocontrol agent from trichoderma harzianum and azadirachtin as claimed in claim 1 for the control of soybean phytophthora blight, characterized in that: the pathogenic bacteria of the soybean phytophthora blight is soybean phytophthoraPhytophthora sojae。
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2019
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|---|---|---|---|---|
| WO2007096833A2 (en) * | 2006-02-27 | 2007-08-30 | North-West University | Composition in the form of a microemulsion containing free fatty acids and/or free fatty acid derivatives |
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