CN111345320A - Application of trichoderma in antagonism of soybean epidemic disease - Google Patents
Application of trichoderma in antagonism of soybean epidemic disease Download PDFInfo
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- CN111345320A CN111345320A CN202010036679.5A CN202010036679A CN111345320A CN 111345320 A CN111345320 A CN 111345320A CN 202010036679 A CN202010036679 A CN 202010036679A CN 111345320 A CN111345320 A CN 111345320A
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Abstract
The invention provides application of trichoderma in antagonism of soybean epidemic disease, and relates to the technical field of biocontrol microbial agents.
Description
Technical Field
The invention belongs to the technical field of biocontrol microbial agents, and particularly relates to application of trichoderma in antagonism of soybean epidemic disease.
Background
In the famous records of quarantine pests of plants in the border of the people's republic of China, southern stem canker of soybean (Diaporthe phaseolorum var. meridionalis) and phomopsis longicolla of soybean are the quarantine pests with the highest interception frequency at the border of soybean ports in China. According to the statistics of customs data, in the soybeans entering the environment in 2006 to date, 230 batches of southern stem canker pathogenic bacteria of the soybeans are intercepted, and 7036 batches of phomopsis longissima are intercepted.
The southern stem canker germ is one of 13 quarantine pathogenic bacteria of soybeans which are of major concern in China, is reported for the first time in the southern part of the United states in 1973, then commonly occurs in various states in the southern part of the United states, and the loss rate is up to 100 percent in areas with serious disasters. The pathogenic bacteria can cause the withering of soybean seedlings, and can cause the death of soybeans in a mature period in severe cases, but cannot cause the influence on human health. At present, the drug is mainly distributed in the countries of the United states, Brazil, Paraguay, Argentina, Viviana and the like. In recent years, epidemic areas are continuously expanded, and China is not distributed at present. Pathogenic bacteria can be spread in a long distance through diseased seeds and diseased residues, and can be spread by port unloading, leakage in the transportation process, screening of leftovers of a grease processing plant and the like. As the soybean production area in China generally has the condition for the survival and growth of the pathogenic bacteria, once the pathogenic bacteria are introduced into China, the pathogenic bacteria bring about great economic loss to the production and planting of the soybeans in China.
The phomopsis longituba is also an important disease in soybean production, can cause soybean root rot and seedling stage diseases, and can reduce germination rate, emergence rate and seed vigor, and can cause that more than 90 percent of seeds can not germinate. Meanwhile, the quality of the soybean seeds is affected, the soybean seeds are faded, and the quality of the soybeans is reduced. It has also been reported that if the disease infection rate with soybeans exceeds 40%, significant yield losses are incurred, which in 1994 results in 18 million tons of soybeans being lost worldwide, with 3.15 million tons being lost in the united states. At present, the disease is reported in the United states, Argentina, Italy, Canada, France, Hungary, Romania, Korea and the like, and no harm report exists in China. In recent years, the disease is widespread abroad, so that great economic loss is caused to soybean crops, and the introduction risk is extremely high.
At present, the two diseases are mainly controlled by medicines. Indoor toxicity experiments show that 50% of carbendazim, 25% of difenoconazole and 5% of hexaconazole can be used as effective bactericides for the two diseases. The benomyl is selected to prevent and treat the soybean phomopsis rot and the soybean stem ulcer in foreign related reports. On one hand, the chemical reagent is adopted, so that pesticide residue is caused, the environment is polluted, the utilization rate of the pesticide is generally 10% according to literature reports, and about 90% of the pesticide residue is left in the environment, so that the environment is polluted; on the other hand, pesticides can be harmful to human health, and cause acute poisoning in a short time after entering human body through respiratory tract, mouth or contact, or cause accumulative poisoning after being eaten by people through gradual enrichment of food chain.
Disclosure of Invention
In view of the above, the present invention aims to provide an application of Trichoderma in antagonizing soybean phytophthora blight, Trichoderma harzianum (Trichoderma harzianum), Trichoderma viride (Trichoderma viride) and/or Trichoderma spiratum (Trichoderma spirale), which have a relatively good antagonistic effect on soybean southern stem canker (Trichoderma longiradiatum) and/or soybean Phomopsis longifola, and can be used as a standard strain for biological control.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides application of Trichoderma in antagonism of soybean phytophthora blight, wherein the Trichoderma comprises Trichoderma harzianum (Trichoderma harzianum), Trichoderma viride (Trichoderma viride) and/or Trichoderma spiraeum (Trichoderma spirale).
Preferably, the Trichoderma harzianum comprises Trichoderma harzianum (Trichoderma harzianum) P9 with the preservation number of CGMCC No. 19267.
Preferably, the Trichoderma viride comprises Trichoderma viride (Trichoderma viride) P8 with the collection number of cgmccno. 19266.
Preferably, the Trichoderma helicatum comprises Trichoderma helicatum (Trichoderma spirale) P10 with the preservation number of CGMCC No. 19268.
Preferably, the soybean quarantine disease includes southern stem canker of soybean (Diaporthe phaseolorum) and/or Phomopsis longifola (Phomopsis longifola).
The invention also provides a soybean epidemic disease detection bacteriostatic agent which is characterized in that the active ingredient of the bacteriostatic agent comprises the trichoderma.
The invention provides application of Trichoderma in antagonism of soybean phytophthora blight, in the embodiment, Trichoderma harzianum (Trichoderma harzianum), Trichoderma viride (Trichoderma viride) and Trichoderma spirans (Trichoderma spirale) are separated and used for performing bacteriostasis loop experiments on Trichoderma sojae (Diaporthe phaseolorum var. meridionalis) and Trichoderma sojae (Phomopsis longicola), and the results show that the Trichoderma harzianum (Trichoderma harzianum), Trichoderma viride (Trichoderma virride) and Trichoderma spirans (Trichoderma spirale) all have good bacteriostasis effect and antagonism on Trichoderma sojae (Trichoderma harzianum var. meridionalis) and Trichoderma sojae (Trichoderma spirans) and can be used as biological control standard strains.
Biological preservation information
Trichoderma harzianum (Trichoderma harzianum) P9, wherein the preservation unit is the China general microbiological culture Collection center, the preservation address is the institute of microbiology, China academy of sciences, No. 3, West Lu 1 Hospital, Kyowa, Beijing, the preservation time is 2019, 12 and 23 days, and the preservation number is CGMCC No. 19267;
trichoderma viride (Trichoderma viride) P8, wherein the preservation unit is China general microbiological culture Collection center, the preservation address is the microbiological research institute of China academy of sciences No. 3, West Lu No.1 Hopkin, Tokyo, sunny district, the preservation time is 12 and 23 days in 2019, and the preservation number is CGMCC No. 19266;
trichoderma spirillum (Trichoderma spirale) P10, the preservation unit is China general microbiological culture Collection center, the preservation address is the microbiological research institute of China academy of sciences No. 3, West Lu No.1 Hopkins, Kyowa, Yangyang, the preservation time is 2019, 12 and 23 days, and the preservation number is CGMCC No. 19268.
Drawings
FIG. 1 is a graph of antagonism of Trichoderma harzianum and southern stem canker of Glycine max, wherein the left graph shows the positive effect of antagonism of two fungi, and the right graph shows the negative effect of antagonism of two fungi;
FIG. 2 is a graph of antagonism of Trichoderma harzianum and Soy bean species Fusarium oxysporum, wherein the left graph shows the positive effect of antagonism of two fungi, and the right graph shows the negative effect of antagonism of two fungi;
FIG. 3 is a graph of antagonism of Trichoderma viride and southern stem canker of Glycine max, wherein the left graph shows the positive effect of antagonism of two fungi, and the right graph shows the negative effect of antagonism of two fungi;
FIG. 4 is a graph of antagonism of Trichoderma viride and Soy bean species Fusarium oxysporum, wherein the left graph shows the positive effect of antagonism of two fungi, and the right graph shows the negative effect of antagonism of two fungi;
FIG. 5 is a graph of antagonism of Trichoderma spiralatum and southern stem canker of Glycine max, wherein the left graph shows the positive effect of antagonism of two fungi, and the right graph shows the negative effect of antagonism of two fungi;
FIG. 6 is a graph showing the antagonistic action of Trichoderma spiralatum and Soy bean species Fusarium, wherein the left side is a graph showing the positive effect of the antagonistic action of the two fungi, and the right side is a graph showing the negative effect of the antagonistic action of the two fungi.
Detailed Description
The invention provides application of Trichoderma in antagonism of soybean phytophthora blight, wherein the Trichoderma comprises Trichoderma harzianum (Trichoderma harzianum), Trichoderma viride (Trichoderma viride) and/or Trichoderma spiraeum (Trichoderma spirale).
The soybean quarantine diseases comprise soybean south stem canker (Amersham Phaseolorvar) and/or soybean Phomopsis longifola.
The trichoderma harzianum is preferably trichoderma harzianum P9 which is preserved with the preservation number of CGMCC No. 19267; the trichoderma viride is preferably trichoderma viride P8 which is preserved with the preservation number of CGMCC No. 19266; the Trichoderma spiralatum is preferably Trichoderma spiralatum P10 with the preservation number of CGMCC No. 19268.
In the invention, an antibacterial zone experiment shows that after Trichoderma harzianum (Trichoderma harzianum) is contacted with southern stem canker pathogenic bacteria (Diaporthe phaseolorum var. meridionalis), the southern stem canker pathogenic bacteria are inhibited, the growth speed is reduced, the contact part has obvious shrinkage traces after 5 days, the contact edge has an obvious isolation zone, and the Trichoderma harzianum forms an enclosure ring for the periphery of the southern stem canker pathogenic bacteria; after the Trichoderma harzianum and the soybean seed rot germ (Phomopsis longicolea) are contacted, the soybean seed rot germ is inhibited, the growth speed is slowed down, the contact part has obvious atrophy traces after 5 days, the contact edge has an obvious isolation zone, and the Trichoderma harzianum forms a surrounding ring for the periphery of the soybean seed rot germ; therefore, trichoderma harzianum has antagonism to southern stem canker and soybean seed rot;
after Trichoderma viride (Trichoderma viride) is contacted with two bacteria of southern stem canker of soybean (diaporthe phaseolorum var. meridionalis), the southern stem canker of soybean is inhibited, the growth speed is slowed down, the contact part has obvious atrophy traces after 5d, the contact edge has obvious isolation zones, and the Trichoderma viride forms a surrounding ring for the periphery of the southern stem canker of soybean; after the Trichoderma viride (Trichoderma viride) and the soybean seed rot fungus (Phomopsis longicola) are contacted, the soybean seed rot fungus is inhibited, the growth speed is slowed down, the contact part has obvious atrophy traces after 5 days, the contact edge has an obvious isolation zone, and the Trichoderma viride forms a surrounding ring for the periphery of the soybean seed rot fungus; therefore, the trichoderma viride has antagonistic action on the southern stem canker and the soybean seed rot germs;
after Trichoderma spirillum (Trichoderma spirale) and southern stem canker of soybean (diaporthe phaseolorum var. meridionalis) are contacted, the southern stem canker of soybean is inhibited, the growth speed is slowed down, the contacted part has obvious atrophy traces after 5d, the contacted edge has obvious isolation zones, and the Trichoderma spirillum forms a surrounding ring for the periphery of the southern stem canker of soybean; after the Trichoderma spirillum and the soybean seed rot (Phomopsis longicola) are contacted, the soybean seed rot is inhibited, the growth speed is slowed down, the contact part has obvious atrophy traces after 5 days, the contact edge has an obvious isolation zone, and the Trichoderma spirillum forms a surrounding ring for the periphery of the soybean seed rot. Therefore, the curly wood has antagonism to southern stem canker and soybean seed rot.
The invention also provides a bacteriostatic agent for detecting the soybean epidemic diseases, and the active component of the bacteriostatic agent comprises the trichoderma. The dosage form of the inhibitor is not particularly limited in the present invention, and conventional dosage forms in the art may be used. The inhibitor of the invention preferably further comprises acceptable auxiliary materials.
The application of trichoderma in antagonizing soybean phytophthora infestans provided by the present invention will be described in detail with reference to the following examples, but they should not be construed as limiting the scope of the present invention.
Example 1
Placing Trichoderma harzianum and southern stem canker of soybean on a PDA culture medium, culturing for 5 days in an incubator at 25 ℃, punching by using a puncher, beating the germs into bacterium blocks with the diameter of about 0.5cm, then respectively picking one bacterium block, placing the bacterium blocks in the same PDA culture dish in a straight line opposite manner at the interval of about 6cm, finally placing the culture dish in the incubator at 25 ℃ for culturing, and observing the growth state of the bacterium colony at intervals of 24 h.
The result is shown in figure 1, after 3 days, the contact between trichoderma harzianum and the two strains of southern stem canker of soybean can be seen, in the later observation, the two strains are inhibited after meeting, the growth speed is reduced, after 5 days, the contact part has obvious atrophy traces, the contact edge has obvious isolation zones, and the trichoderma harzianum forms a surrounding ring for the periphery of the southern stem canker of soybean. Thus, trichoderma harzianum has an antagonistic effect on southern stem canker of soybean.
Example 2
Placing Trichoderma harzianum and soybean seed rot bacteria on a PDA culture medium, culturing for 5 days in an incubator at 25 ℃, punching by using a puncher, beating the bacteria into bacterial blocks with the diameter of about 0.5cm, then respectively selecting one bacterial block, placing the bacterial blocks in the same PDA culture dish in a straight line opposite manner at the interval of about 6cm, finally placing the culture dish in the incubator at 25 ℃ for culturing, and observing the growth state of bacterial colonies every 24 hours.
As shown in the figure 2, after 3 days, the contact between the two strains of Trichoderma harzianum and the soybean species pythium is seen, and in the later observation, the two strains are inhibited after meeting each other, the growth rate is reduced, after 5 days, the contact part has obvious atrophy traces, the contact edge has obvious isolation zones, and the periphery of the soybean species pythium is surrounded by the Trichoderma harzianum. Therefore, trichoderma harzianum has antagonistic action on soybean seed rot.
Example 3
The method comprises the steps of placing trichoderma viride and soybean southern stem canker pathogenic bacteria on a PDA culture medium, culturing for 5 days in an incubator at 25 ℃, punching by using a puncher, beating the pathogenic bacteria into bacterial blocks with the diameter of about 0.5cm, then respectively picking one bacterial block, placing the bacterial blocks in the same PDA culture dish in a straight line opposite mode with the distance of about 6cm, finally placing the culture dish in the incubator at 25 ℃ for culturing, and observing the growth state of bacterial colonies every 24 hours.
The result is shown in fig. 3, after 3 days, the trichoderma viride and the southern stem canker of soybean can be seen to be contacted, and in the later observation, the two strains are inhibited after meeting each other, the growth speed is reduced, after 5 days, the contact part has obvious atrophy traces, the contact edge has obvious isolation zones, and the trichoderma viride forms a surrounding ring for the periphery of the southern stem canker of soybean. Therefore, trichoderma viride has an antagonistic effect on southern stem canker of soybean.
Example 4
Placing Trichoderma viride and soybean seed rot bacteria on a PDA culture medium, culturing for 5 days in an incubator at 25 ℃, punching by using a puncher, beating the bacteria into bacterial blocks with the diameter of about 0.5cm, then respectively selecting one bacterial block, placing the bacterial blocks in the same PDA culture dish in a straight line opposite manner at the interval of about 6cm, finally placing the culture dish in the incubator at 25 ℃ for culturing, and observing the growth state of bacterial colonies every 24 hours.
As shown in FIG. 4, after 3 days, the contact between Trichoderma viride and Soy bean species Humicola was observed, and in the subsequent observation, the two strains were inhibited after meeting each other, the growth rate was reduced, after 5 days, the contact part had obvious atrophy trace, the contact edge had obvious isolation zone, and Trichoderma viride formed a surrounding ring around Soy bean species Humicola. Therefore, the trichoderma viride has an antagonistic effect on soybean seed rot.
Example 5
Placing Trichoderma spirillum and southern stem canker pathogenic bacteria on a PDA culture medium, culturing for 5 days in an incubator at 25 ℃, punching by using a puncher, beating the pathogenic bacteria into bacterial blocks with the diameter of about 0.5cm, then respectively picking one bacterial block, placing the bacterial blocks in the same PDA culture dish in a straight line opposite manner with the distance of about 6cm, finally placing the culture dish in the incubator at 25 ℃ for culturing, and observing the growth state of bacterial colonies every 24 hours.
The result is shown in fig. 5, after 3 days, the contact between the two strains of the trichoderma spiratum and the southern stem canker of the soybean can be seen, in the later observation, the two strains are inhibited after meeting, the growth speed is reduced, after 5 days, the contact part has obvious atrophy traces, the contact edge has obvious isolation zones, and the trichoderma spiratum forms a surrounding ring for the southern stem canker of the soybean. Therefore, trichoderma spirillum has antagonistic effect on southern stem canker of soybean.
Example 6
Placing Trichoderma spirillum and Soy bean species pythium in a PDA culture medium, culturing for 5 days in an incubator at 25 ℃, punching by using a puncher, beating the germs into bacterial blocks with the diameter of about 0.5cm, then respectively selecting one bacterial block, placing the bacterial blocks in the same PDA culture dish in a straight line opposite way at the interval of about 6cm, finally placing the culture dish in the incubator at 25 ℃ for culturing, and observing the growth state of bacterial colonies every 24 hours.
As shown in FIG. 6, after 3 days, the contact between the two strains of Trichoderma spiralatum and Soy bean species Humicola was observed, and in the subsequent observation, the two strains were inhibited after meeting each other, the growth rate was reduced, after 5 days, the contact part had obvious atrophy trace, the contact edge had obvious isolation zone, and the Trichoderma spiralatum formed a surrounding ring for the periphery of Soy bean species Humicola. Therefore, the trichoderma spirillum has antagonistic effect on soybean seed rot.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (6)
1. The application of Trichoderma in antagonizing soybean phytophthora blight is characterized in that the Trichoderma comprises Trichoderma harzianum (Trichoderma harzianum), Trichoderma viride (Trichoderma viride) and/or Trichoderma spiraeum (Trichoderma spirale).
2. The use according to claim 1, wherein the Trichoderma harzianum comprises Trichoderma harzianum (Trichoderma harzianum) P9, accession No. CGMCC No. 19267.
3. The use according to claim 1, wherein the Trichoderma viride strain comprises Trichoderma viride (Trichoderma viride) P8, accession No. CGMCC No. 19266.
4. The use according to claim 1, wherein the Trichoderma helicidum comprises Trichoderma helicidum (Trichoderma spirale) P10, accession No. CGMCC No. 19268.
5. The use of claim 1, wherein said soybean quarantine disease includes southern stem canker (Diaporthephaeroleum var. meridionalis) and/or phomopsis longicolla (Phomopsis longicolla).
6. A soybean quarantine pest bacteriostat, wherein the active ingredient of the bacteriostat comprises the trichoderma in the application of claim 1.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN114958613A (en) * | 2022-04-02 | 2022-08-30 | 四川农业大学 | Trichoderma spirillum RS05 and application thereof in preventing and treating brown rot of dendrocalamus latiflorus |
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| CN102823467A (en) * | 2012-09-21 | 2012-12-19 | 农业部环境保护科研监测所 | Method for controlling legume root rot by using trichoderma viride |
| WO2017173689A1 (en) * | 2016-04-07 | 2017-10-12 | 吴政宏 | Application using tolypocladium inflatum to manufacture shrimp feed and/or formulation for treating or preventing vibriosis |
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| CN102823467A (en) * | 2012-09-21 | 2012-12-19 | 农业部环境保护科研监测所 | Method for controlling legume root rot by using trichoderma viride |
| WO2017173689A1 (en) * | 2016-04-07 | 2017-10-12 | 吴政宏 | Application using tolypocladium inflatum to manufacture shrimp feed and/or formulation for treating or preventing vibriosis |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN114958613A (en) * | 2022-04-02 | 2022-08-30 | 四川农业大学 | Trichoderma spirillum RS05 and application thereof in preventing and treating brown rot of dendrocalamus latiflorus |
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