CN116491524B - Application and method for controlling mulberry sclerotinia by using coniothyrium minitans - Google Patents
Application and method for controlling mulberry sclerotinia by using coniothyrium minitans Download PDFInfo
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Classifications
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G13/00—Protecting plants
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G17/00—Cultivation of hops, vines, fruit trees, or like trees
- A01G17/005—Cultivation methods
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
- A01P3/00—Fungicides
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Zoology (AREA)
- Microbiology (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Mycology (AREA)
- Wood Science & Technology (AREA)
- Pest Control & Pesticides (AREA)
- Plant Pathology (AREA)
- Virology (AREA)
- Dentistry (AREA)
- Biotechnology (AREA)
- Agronomy & Crop Science (AREA)
- Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
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- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention relates to the technical field of microbial pesticides, in particular to application and a method for controlling mulberry sclerotinia by using coniothyrium minitans. The application of the invention has better control effect by using the coniothyrium minitans for controlling the mulberry sclerotinia rot. The method of the invention only needs to spray the preparation containing the coniothyrium minitans after pruning branches of the mulberry in winter or when the mulberry germinates, and the invention has the advantages of simplicity, convenience and long acting time, and the control effect is equivalent to the effect of repeatedly spraying pesticides in the prior art.
Description
Technical Field
The invention relates to the technical field of microbial pesticides, in particular to application and a method for controlling mulberry sclerotinia by using coniothyrium minitans.
Background
The mulberry sclerotinia is also called a ginkgo disease, is a main disease of mulberry, can cause no harvest of mulberry particles when the damage is serious, and causes pathogenic spores to spread along with air flow (with high speed and wide range), so that female flowers and green fruits are infected, new shoots and tender buds of early-growing mulberry are mainly infected, after the female flowers are invaded, pulp swells, the pulp is milky white and grey brown, and rotten odor can be smelled after the twisting.
Sclerotinia sclerotiorum is a worldwide broad-spectrum parasitic pathogenic fungus, and the purpose of radically treating the disease cannot be achieved by adopting chemical measures. In order to reduce pesticide residues and avoid the enhancement of pathogen resistance, people are hoped to be supported in the aspects of development and utilization of biocontrol agents. Among a plurality of microbial flora with biological inhibition effect on sclerotinia sclerotiorum, the coniothyrium minitans is the microorganism with the most development potential for preventing and treating sclerotinia sclerotiorum at present, can selectively parasitize sclerotinia sclerotiorum formed by the sclerotinia sclerotiorum, can effectively and exclusively decompose overwintering sclerotium of pathogenic bacteria in soil and inhibit germination of the sclerotinia sclerotiorum, and can survive in the soil for a plurality of years, and the microorganism has the characteristics of strong specificity, long acting time, no pathogenicity to animals and plants and the like, and is an ideal biological prevention and treatment agent with the most development potential for preventing and treating sclerotinia sclerotiorum. Currently, the coniothyrium minitans is mainly used for preventing and treating rape and vegetable sclerotinia, and is not reported for preventing and treating the mulberry sclerotinia.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provides an application and a method for controlling mulberry sclerotinia by using coniothyrium minitans.
The aim of the invention is realized by the following technical scheme:
The invention provides an application of coniothyrium minitans in preventing and treating mulberry sclerotinia. The coniothyrium minitans has a good effect by using the coniothyrium minitans for preventing and treating the mulberry sclerotinia.
The disease origin of the mulberry sclerotinia is different from the pathogen of sclerotinia such as rape, vegetables and the like. The pathogen of sclerotinia rot of rape and vegetables is sclerotinia sclerotiorum (Sclerotinia sclerotiorum (Lib.) de Bary), belongs to ascomycota, sclerotiniaceae, sclerotinia fungus, and sclerotinia sclerotiorum can generate three forms of hyphae, sclerotium and ascospore under natural conditions, the sclerotium is used for invading and infecting hosts by the hyphae, but the main form is sclerotium, and sclerotium can wait for proper growth environment in soil or plant residues to germinate and grow. The sclerotinia sclerotiorum hosts are very wide, can parasitize more than 400 monocotyledonous and dicotyledonous plants worldwide, and the infection and prevention of sclerotinia sclerotiorum which is an important economic crop such as rape, soybean, peanut and the like are more studied at present. Mulberry hypertrophic sclerotinia bacteria Ciboria shiraiana P.Henn. Called Mulberry sclerotiorum (Leuconostoc mesenteroides), belongs to fungus of genus Leuconostoc of sub-phylum ascomycotina, and Mulberry sclerotium is difficult to culture in culture medium, which is one of the main difficulties in researching the fungus, R.Sultana and K.Kim. (2016) consider that Mulberry sclerotinia is difficult to grow on artificial PDA or other culture medium, and the characteristic that Mulberry hypertrophic sclerotinia fungus may be the fungus of specific parasitic mulberry. The pathogenic bacteria overwintere in soil with sclerotium, the sclerotium falls to the ground along with the disease, when the female flowers of the next year of mulberry are opened, the sclerotium which has passed through winter in the soil sprouts out ascus discs, and each sclerotium can sprout out one or a plurality of ascus discs. The ascus disk is disk-shaped or cup-shaped, the meat quality is brown, ascus and ascus spores are layered on the ascus disk, after the fruiting body is mature, the ascus spores are ejected into the air to spread along with wind, when the ascus spores fall on female flowers of mulberry, hyphae are germinated to infect the female flowers of the mulberry, and the occurrence of the sclerotium disease of the mulberry is caused. Mulberry miniprep. Scleromitrula shiraiana (henn.) S.Ima called Polygonum nucleatum, also called Bai Jing Polygonum nucleatum (Mitrula shiraiana (P.henn.) Ito etlmai), belongs to the phylum ascomycota, is a species of Mulberry-gingko disease pathogen that can be easily isolated and grown on artificial media, but so far no report has been made on the finding that other plants are infested with the Polygonum nucleatum, thus deducing that the Polygonum nucleatum may still be a specific pathogen that only parasitizes Mulberry but is not pathogenic to other plants. Mulberry sclerotinia sclerotiorum Ciboria carunculoides SIEGLER ET Jankins is called calicheapest, belonging to the genus calicheapest of the family Coptidaceae, the subglotriopsis of the genus Coptis. Overwintering in soil in the form of sclerotium, starting germination of fruiting bodies when the soil is warmed up by weather and is wet by rainwater, erupting ascospores, and growing along the flower organ of mulberry when the ascospores fall on a proper host by air flow, so that infection of small mulberry ovaries is caused. The isolated culture of calixania carnis is difficult, and so far there are few reports of in vitro culture, indicating that it is possible to have a specific infection Sang Cihua.
The mechanism of the coniothyrium minitans for preventing and controlling crop sclerotinia mainly has two aspects of re-mailing effect and antibiotic effect. The successful parasitic infection of sclerotium by coniothyrium minitans, the inhibition of sclerotinia disease, is the result of the combined action of the two mechanisms. Wherein the antibiotic action mainly inhibits the occurrence of diseases by producing antibiotic substances and inhibiting the growth of pathogenic bacterial hyphae. The coniothyrium minitans secretes a series of chitinase, beta-1, 3-glucanase and other substances to lyse the sclerotium cell wall, lay a foundation for hypha infection sclerotium, prevent sclerotium germination and even death, and reduce primary infection sources.
The invention also provides a method for controlling the mulberry sclerotinia by the coniothyrium minitans, which comprises the following steps: spraying a preparation containing the conifer when the mulberry prunes branches in winter or the mulberry germinates.
Considering that the main spreading and popular modes of mulberry ginkgo disease are: the pathogenic bacteria overwintere in soil or plant residues in a sclerotium form, when the climate conditions are proper in the next year, the sclerotium germinates to grow an ascomycete disc, the ascomycete disc matures and ejects to release ascospores, the ascospores are spread and attached to mulberry leaves, petals or mulberries along with wind, and the ascospores germinate and infect again to cause diseases. The sclerotium disease prevention and control of the conidium disease mainly acts from two aspects, namely, the sclerotium is sent to decay by re-mailing, so that the sclerotium cannot germinate to generate ascospores under proper climatic conditions, namely, the primary infection source of the sclerotium disease is reduced, and the disease prevention effect is achieved; secondly, antibiotic substances are generated, hypha growth is inhibited, growth is interrupted, and infection is prevented from reaching the disease control effect. Therefore, the application of the conifer is carried out before sclerotium germination and after mulberry pruning in winter until mulberry germination; secondly, because sclerotium exists in soil or plant residue, after the mulberry is pruned in winter or the mulberry is germinated, the mulberry does not grow mulberry leaves, and the pesticide is sprayed onto the soil without mulberry She Zudang, so that the pesticide can be fully contacted with the sclerotium, and the rotting rate and the disease prevention effect are improved.
Preferably, the spraying of the preparation is a plurality of sprays, and the spraying is performed again after the first spraying of the preparation at intervals of 5-9 days.
Preferably, the specific method for spraying is as follows: spraying the ground below the mulberry and the branches of the mulberry by a spraying mode.
Preferably, the content of conidium molecules in the preparation is 1X 10 9-2×109/g, and the dosage of the preparation is 20-25 g/mu.
Preferably, the formulation is sprayed after mixing with water, a carbon source.
Preferably, the mass ratio of the preparation to the water to the carbon source is as follows: 20-25:40000-50000:8-10.
Preferably, the carbon source is sucrose.
The coniothyrium minitans is a obligate re-parasitic fungus, and has low field survival capability under the conditions of existence of sterile cores in the field and poor nutrition, so that sucrose needs to be added to provide partial carbon source for the growth of the coniothyrium minitans, and the field inventory colonization capability of the coniothyrium minitans is improved. The proper growth temperature of the coniothyrium minitans is 20 ℃, and under the temperature condition, most pathogenic bacteria are in a dormant state, and the addition concentration is extremely low, so that the probability of pathogen growth is low. Thus adding sucrose is more advantageous than the disadvantages. Compared with various nutrient substances such as sucrose, glucose, starch and the like, the technology is applied to the masses at last, and the sucrose is relatively more convenient to purchase by combining an indoor coniothyrium minitans carbon source selection and utilization experiment, so that the sucrose is recommended to be used as an additive substance.
Preferably, the mulberries sclerotinia is caused by one or more of Bai Jing cane fungus, leucotrichum candidum and leucotrichum mongolicum.
Preferably, the storage number of the coniothyrium minitans is: CGMCC N0.3852.
The beneficial effects of the invention are as follows:
1. The invention provides an application of coniothyrium minitans in preventing and treating the sclerotinia sclerotiorum, which has a good preventing and treating effect by using coniothyrium minitans in preventing and treating the sclerotinia sclerotiorum.
2. The invention provides a method for preventing and controlling mulberry sclerotinia by using coniothyrium minitans, which is simple, convenient and long in acting time, and has the prevention and control effect equivalent to the effect of repeatedly spraying pesticides in the prior art, and a preparation containing the coniothyrium minitans is sprayed after the branches of the mulberry are pruned or the mulberry germinates in winter.
Drawings
FIG. 1 shows the antagonism of the culture of the coniothyrium minitans Cm2004 against the pathogenic bacteria of the sclerotinia sclerotiorum;
Fig. 2 shows that the untreated mulberry of the coniothyrium minitans Cm2004 preparation is suffering from the ginkgo disease;
Fig. 3 is a view of a coniothyrium minitans Cm2004 formulation treating mulberry.
Detailed Description
The technical solution of the present invention will be described in further detail with reference to the accompanying drawings, but the scope of the present invention is not limited to the following description.
EXAMPLE 1 Effect of Thermomyces lanuginosus on inhibition of Leuconostoc mesenteroides, bai Jing Di Zhang Jun and Leuconostoc morchella
(1) Experimental conditions
According to the invention, two different coniothyrium minitans are selected, and an experiment is carried out by using bacillus amyloliquefaciens, wherein the experimental conditions are as follows:
(2) Experimental methods and results
The 3 different strains are used for opposite culture with pathogenic bacteria of mulberry and ginkgo, clear water is used as a blank control for opposite culture, the colony diameter of the pathogenic bacteria is measured after 20 days, and the growth inhibition rate is converted, and the results are shown in fig. 1 and table 1:
TABLE 1 comparison of growth inhibition effect (%) of biocontrol strain on Mulberry Ginkgo germ
| Strain | Cm2004 | Bam22 | Cm315 |
| Brevibacterium leucotrichum | 88.25 | 84.65 | 83.23 |
| Bai Jing Polygonum cuspidatum | 86.23 | 80.54 | 80.14 |
| Calixarene of caruncle | 87.24 | 81.54 | 80.27 |
Bacillus amyloliquefaciens is a known strain with disease resistance, bacteriostasis and broad-spectrum high efficiency, and the antibacterial effect of the coniothyrium minitans used by the invention is equivalent to that of the Bacillus amyloliquefaciens, and especially the antibacterial effect of Cm2004 is even better than that of the Bacillus amyloliquefaciens. Shows that the coniothyrium minitans has better potential for preventing and treating the mulberry sclerotinia.
EXAMPLE 2 Effect of different carbon sources on the growth of Phytophthora
Different carbon sources are adopted and respectively added into a conidium fermenting matrix, the mass ratio of the carbon sources is 10:1, conidium is cultivated in darkness at 20 ℃ for 15-20 days, the influence of the different carbon sources on conidium growth of the conidium is explored, and the experimental data are obtained by eluting after the cultivation is finished and counting by a blood cell counting plate, wherein the experimental data are as follows:
TABLE 3 Effect of carbon source difference on growth of coniothyrium minitans
| Carbon source | Sucrose | Glucose | Maltose | Starch |
| Spore yield (individual/g) | 2.3×109 | 1.2×109 | 9.7×108 | 9.2×108 |
Example 3 comparative experiment of the amount of Shell mould
Selecting the molecular spore content of the conidium preparation of about 1X 10 9-2×109/g, and spraying 50 liters of water for medicine/667 m 2; when 10g/667m 2 of sucrose is added, different application quality grade difference treatment tests are set, and the control effect of the use amount of the coniothyrium minitans on the mulberry sclerotinia is shown in Table 2:
TABLE 2 influence of the amount of Phyllostachys pubescens on the control effect of Mulberry sclerotiorum
Example 4 comparison of effects of Phytophthora on controlling Mulberry sclerotiorum
(1) Experimental conditions and methods
Treatment area: after pruning branches of the mulberry in winter or in the germination period of the mulberry, spraying the conifer preparation twice, wherein the spraying interval period is 7 days, and the two spraying modes are the same; when spraying, the ground soil is mainly sprayed and the mulberry branches are properly sprayed. 3 replicates were set.
The coniothyrium minitans in the coniothyrium minitans preparation is coniothyrium minitans (Coniothyrium minitans) Cm2004, and is preserved in China general microbiological culture Collection center (China Committee) with the preservation number: CGMCC N0.3852. The amount of the coniothyrium minitans in the coniothyrium minitans preparation is as follows: the molecular spore content is about 1X 10 9-2×109/g, sucrose and water are additionally added, the dosage per mu is 20-25g of conidium, 40-50L of water and 8-10g of sucrose.
Positive control: the chemical agent is sprayed in real time in a motorized way in the growing period, and the ground, the branches and the leaves are sprayed during the spraying process, so that dead angles are not remained. 3 replicates were set. The method comprises the following steps: during the germination period of the mulberry, 50% Kazier water dispersible granule (24 g/667m 2, 50L water/667 m 2) is sprayed once. The mulberry is treated twice in the vigorous growth period, and the treatment interval is 6-10 days; the first medicament is 400 times liquid spray treatment of 80% Dasheng-M45 wettable powder; the second application agent is 50% Kazier water dispersible granule, and the treatment mode is the same as that of the mulberry in the germination period.
Negative control: no treatment with any agent. Three treatments were arranged in a random block.
(2) Experimental results
The mature period of the mulberries is spotted according to a quincuncial pattern, and representative 5 plants are randomly extracted from each area to survey the number of the mulberries with sclerotium disease, the fruit yield and the converted theoretical fruit yield of 667 square meters, wherein the theoretical fruit yield is shown in the following table 1:
TABLE 1
As can be seen from the table, compared with the negative control, the method for controlling the mulberry sclerotinia by using the scutellum provided by the invention has the advantages that the fruit yield is obviously improved, the gingko quantity is reduced, and the effect of controlling the mulberry sclerotinia is effectively achieved; compared with the positive control, although the effect of the positive control is slightly inferior, the effect of the scutellum preparation is more than that of the positive control after the effective components in the scutellum preparation are optimized by using only two effective components of scutellum and sucrose; shows that the coniothyrium minitans has great potential in preventing and treating the mulberry sclerotinia.
The foregoing is merely a preferred embodiment of the invention, and it is to be understood that the invention is not limited to the form disclosed herein but is not to be construed as excluding other embodiments, but is capable of numerous other combinations, modifications and environments and is capable of modifications within the scope of the inventive concept, either as taught or as a matter of routine skill or knowledge in the relevant art. And that modifications and variations which do not depart from the spirit and scope of the invention are intended to be within the scope of the appended claims.
Claims (4)
1. Application of coniothyrium minitans (Coniothyrium minitans) Cm2004 in preventing and treating mulberry sclerotinia disease is characterized in that: the scutellum Cm2004 is preserved in China general microbiological culture Collection center (China Committee for culture Collection), and the preservation number is: CGMCC N0.3852; the mulberry sclerotinia is caused by one or more of Bai Jing cane fungus (Mitrula shiraiana (p.henn.) Ito etlmai), leucotrichum (Ciboria shiraiana p.henn.) and leucotrichum caryophyllum (Ciboria carunculoidesSiegler et Jankins).
2. The method for preventing and treating the sclerotinia of mulberries by using the conidium (Coniothyrium minitans) Cm2004 is characterized by comprising the following steps of: the scutellum Cm2004 is preserved in China general microbiological culture Collection center (China Committee for culture Collection), and the preservation number is: CGMCC N0.3852; the mulberry sclerotinia is caused by one or more of Bai Jing cane fungus (Mitrula shiraiana (p.henn.) Ito etlmai), cupula (Ciboria shiraiana p.henn.) and cupula sarcodictyophyllata (Ciboria carunculoidesSiegler et Jankins);
The method comprises the following steps:
Spraying a preparation containing the conifer Cm2004 after pruning branches of the mulberry in winter or when the mulberry germinates; wherein the conidium content of the conidium of the preparation is 1 multiplied by 10 9-2×109 per gram, and the dosage of the preparation is 20-25g per mu; the preparation is sprayed after being mixed with water and a carbon source; the mass ratio of the preparation to the water to the carbon source is as follows: 20-25:40000-50000:8-10; the carbon source is sucrose.
3. The method according to claim 2, characterized in that: spraying the preparation for multiple times, and spraying again after 5-9 days after the first spraying.
4. The method according to claim 2, characterized in that: the specific method for spraying comprises the following steps: spraying the ground below the mulberry and the branches of the mulberry by a spraying mode.
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| CN101603009A (en) * | 2009-06-05 | 2009-12-16 | 华中农业大学 | One strain prevents and treats the mould ZS-1SB of biocontrol microorganisms shield shell and the preparation method and the application of sclerotium disease |
| CN102234619A (en) * | 2011-06-16 | 2011-11-09 | 四川省农业科学院植物保护研究所 | Biocontrol bacteria coniothyrium minitans Cm2004 strain for preventing and treating sclerotiniose and preparation method and application thereof |
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| KR20090011139A (en) * | 2007-07-25 | 2009-02-02 | 대한민국(관리부서:농촌진흥청장) | Coniothyrium minitans cm2 and methods of controlling pathogenic plant fungi by using the same |
| US9788551B2 (en) * | 2011-05-27 | 2017-10-17 | Bayer CropScience Biologies GmbH | Liquid preparation for biological plant protection, method for producing it and use thereof |
| WO2018128985A1 (en) * | 2017-01-03 | 2018-07-12 | Monsanto Technology Llc | Microbial compositions and methods |
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| CN101603009A (en) * | 2009-06-05 | 2009-12-16 | 华中农业大学 | One strain prevents and treats the mould ZS-1SB of biocontrol microorganisms shield shell and the preparation method and the application of sclerotium disease |
| CN102234619A (en) * | 2011-06-16 | 2011-11-09 | 四川省农业科学院植物保护研究所 | Biocontrol bacteria coniothyrium minitans Cm2004 strain for preventing and treating sclerotiniose and preparation method and application thereof |
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