CN111471598A - Gliocladium roseum and pimavalia rimonaris composite microbial agent and application thereof in prevention and control of diseases and pests - Google Patents
Gliocladium roseum and pimavalia rimonaris composite microbial agent and application thereof in prevention and control of diseases and pests Download PDFInfo
- Publication number
- CN111471598A CN111471598A CN202010336121.9A CN202010336121A CN111471598A CN 111471598 A CN111471598 A CN 111471598A CN 202010336121 A CN202010336121 A CN 202010336121A CN 111471598 A CN111471598 A CN 111471598A
- Authority
- CN
- China
- Prior art keywords
- gliocladium roseum
- microbial inoculum
- composite microbial
- fusarium
- gliocladium
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Microbiology (AREA)
- Virology (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Environmental Sciences (AREA)
- Botany (AREA)
- Mycology (AREA)
- Agronomy & Crop Science (AREA)
- Medicinal Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Pest Control & Pesticides (AREA)
- Biomedical Technology (AREA)
- Dentistry (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Plant Pathology (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention relates to the technical field of microorganisms, in particular to a gliocladium roseum and nemoras terrestris composite microbial agent and application of the gliocladium roseum and the nemoras terrestris composite microbial agent in disease control. The Gliocladium roseum is Gliocladium roseum (Clinosstachysolea) fhnzm, the preservation number is CGMCC No.17070, the Monilia similis is Muslim mosaic (Massiitiamoniae) BMMSJS-1, and the preservation number is CGMCC No. 18668. The compound microbial inoculum bacterial liquid can fix nitrogen, promote the growth of seedlings, preferentially symbiotic with the seedlings in a sterile soilless substrate seedling raising mode, and breed high-quality seedlings carrying probiotics. The effect of the composite microbial inoculum bacterial liquid on the growth promotion and the pest and disease protection of the alpine shade plants is better than that of probiotic bacteria. The composite microbial inoculum bacterial liquid can be used for preventing and treating root rot, rough mouth disease and the like of mountain shade plants and promoting the growth of mountain nitrogen-rich plants.
Description
Technical Field
The invention relates to the technical field of microorganisms, in particular to a gliocladium roseum and pimelia rimosus compound microbial inoculum and application of the compound microbial inoculum in prevention and treatment of diseases and pests.
Background
Fusarium (Fusarium L ink ex Fr), also called Fusarium, is taxonomically a plant of the genus Fusarium which originally belongs to deuteromycotina at the asexual stage and belongs to the order of hymenophora, and is ascomycetina at the sexual stage, and is often of the genus Gibberella at the sexual state, Fusarium is a worldwide fungus which can not only overwinter and overwinter in soil, but also infect various economic crops to cause various diseases of root rot, stem rot, flower rot and ear rot of plants, and the main diseases are expressed as rot, blight and corn rot, and the like, and host plants are various in various types including medicinal plants, economic crops and the like, which mainly cause crop wilt death, affect yield and quality, are one of the most important diseases for production control.
Gliocladium roseum (Clinostasysrosea) is a widespread heavy-parasitic bacterium, has the advantages of rapid growth, large sporulation quantity, wide host range, strong parasitism, various antagonistic mechanisms and the like, and the antagonistic mechanism mainly comprises heavy-parasitic action, antagonistic action, induced plant resistance and the like, can produce 3 propagules in a growth cycle, comprises mycelium, conidiospore and chlamydospore, and is one of the most potential plant disease biocontrol bacteria in antagonistic microorganisms found at present.
Generally speaking, the embryo in the healthy seed is sterile, after the seed germinates in a sterile matrix, the exogenous bacteria have the opportunity of invasive field planting, at the moment, the artificial probiotic addition can be preferentially field planting, occupies a corresponding ecological niche, and has a competitive advantage of being firstly introduced compared with a 'later person'. In recent years, with the development of organic planting and ecological planting of traditional Chinese medicinal materials, chemically synthesized insecticides and bacteriostats are forbidden, and microbial pesticides and plant pesticides are vigorously developed. However, the practical application effect of the microbial pesticide is influenced by factors such as unstable application effect of the microbial pesticide, adaptability of probiotic microorganisms to the environment, effective colonization of the probiotic microorganisms in soil, interaction with indigenous microorganisms, influence of chemical insecticidal and bactericidal agents remaining in farmland soil on the probiotic microorganisms, and the like. The invention discloses an endophytic fungus of sinopodophyllum hexandrum and application thereof (CN201910282640.9), and discloses a sinopodophyllum hexandrum endophytic fungus, namely gliocladium roseum fhnzm with the preservation number of CGMCC No.17070, which is preserved in the China general microbiological culture collection center. The strain can improve the content of podophyllotoxin in the podophyllum hexandrum root, inhibit the growth of the podophyllum hexandrum root, inhibit the germination of plant seeds and the elongation of radicles, can be used as a core strain to produce a microbial inoculum for promoting the accumulation of the podophyllotoxin, and has great significance for green prevention and control of diseases and insect pests of the podophyllum hexandrum and improvement of the quality of artificially cultured podophyllum hexandrum, and effective improvement of the accumulation of the podophyllotoxin. However, because fungi and bacteria have a universal cross-border antagonistic effect, the study on the gliocladium roseum symbiotic system bacteria is not carried out at present, and on the basis of the study of a patent (CN201910282640.9), the invention unexpectedly obtains the gliocladium roseum symbiotic bacteria which has very high enzyme activity, has a growth promoting effect on gliocladium roseum, can help the gliocladium roseum to inhibit the growth of fusarium pathogenic bacteria and has the effect of preventing and treating plant root rot and blumeria caused by fusarium pathogenic bacteria.
Disclosure of Invention
An object of the present invention is to provide a complex microbial inoculum consisting of Gliocladium roseum and Gliocladium rim.
Preferably, the gliocladium roseum is gliocladium roseum (Clonostachys rosea) fhnzm, is preserved in the China general microbiological culture collection center in 2019, 1 and 14 days, the preservation number is CGMCC No.17070, the preservation address is No. 3 of Western No.1 Hospital No.1 of sunward area in Beijing, the survival state is survival, and the telephone: 010-: 010-64807288.
Preferably, the mortierella morganii is mortierella morganii (Massilia timonae) BMMSJS-1, which is preserved in the general microbial strain preservation management center of china 10 and 11 days in 2019, the preservation number is CGMCC No.18668, the preservation address is No. 3 of west road No.1 north chen of sunny district in beijing, the survival state is survival, and the telephone: 010-: 010-64807288.
The composite microbial inoculum is applied to inhibiting fusarium pathogenic bacteria.
Preferably, the fusarium pathogenic bacteria are fusarium solani or fusarium oxysporum.
The composite microbial inoculum is applied to preventing and treating plant diseases caused by fusarium pathogenic bacteria.
Preferably, the plant is a medicinal plant, and the disease comprises one or more of rot, blight and numb mouth disease caused by fusarium pathogenic bacteria.
Preferably, the diseases comprise fritillaria root rot, angelica root rot and angelica nummularia.
A pesticide for preventing and treating plant diseases caused by fusarium pathogenic bacteria contains a composite microbial inoculum consisting of the gliocladium roseum and the symbiotic fungus thereof, namely the marcescens mosaici.
Preferably, the bacterial count or the spore count of the gliocladium roseum and the tesserella intermedia composite microbial inoculum is 106～107/ml。
The invention has the beneficial effects that: the composite microbial inoculum bacterial liquid claimed by the invention can fix nitrogen, promote the growth of seedlings, preferentially symbiotic with the seedlings in a sterile soilless substrate seedling raising mode under facility conditions, and breed high-quality seedlings carrying probiotics. In the field cultivation stage, the spread way of pathogenic fungi can be cut off by means of seedling coating and root irrigation after cultivation, the pathogenic fungi can be inhibited and killed, and the nematodes and underground pests can be inhibited and killed. The fungus gliocladium roseum in the compound microbial inoculum liquid has the functions of inhibiting and killing bacteria, fungi, nematodes and underground pests, has the main action mechanisms of heavy parasitism and secretion of antagonistic substances, can be propagated in an application environment through spores, and has better effect on the growth promotion of alpine shade plants and the prevention of plant diseases and insect pests than probiotic bacteria. The composite microbial inoculum bacterial liquid can be used for preventing and treating diseases such as root rot and pockmark disease of the alpine shade plants and promoting the growth of the alpine nitrogen-rich plants.
Drawings
FIG. 1 incidence of fritillary bulb seedball in different groups
Detailed Description
The following examples further illustrate the present invention but are not to be construed as limiting the invention and all modifications or alterations to the methods, procedures or conditions of the present invention which fall within the scope of the invention are to be considered as within the spirit and scope of the invention.
The seedling breeding used in the following experiments was from the biological institute of academy of sciences of Gansu province and the greenhouse of pilot plant base (altitude 1800 m). The cultivation test of the simulated environment of the fritillaria kansuensis is located in the county of elm, namely the Ma Pop village (the elevation is 2600 meters), and the cultivation test of domestication is located in the city of Huating, namely the Maxia town (the elevation is 1800 meters). The angelica sinensis cultivation test is located in Min county Shi town (altitude 2200 meters). The field test is a continuous cropping field. The reagents and culture medium used in the test are all chemically pure.
Example I Effect of Gliocladium roseum fhnzm and Thymus moniliforme BMMSJS-1 Complex microbial inoculum on prevention and treatment of potted fritillary root rot
In order to evaluate the prevention and treatment effect of the composite bacterial liquid of gliocladium roseum fhnzm and gliocladium benthicum BMMSJS-1 on the root rot of fritillaria caused by fusarium pathogenic bacteria, the embodiment firstly uses potted fritillaria as an experimental object to carry out an evaluation experiment, the embodiment is divided into an experimental group, a control group 1, a positive control group and a blank group, the experimental group is irrigated with the composite bacterial liquid of gliocladium roseum fhnzm and gliocladium benthicum BMMSJS-1, the control group 1 is irrigated with the composite bacterial liquid of gliocladium roseum fhnzm, the positive control group is irrigated with the bacillus amyloliquefaciens, the blank group is irrigated with sterilizing water, and the irrigated bacterial liquid is the cultured original bacterial liquid (the bacterial number or the spore number is 106-107Per ml), diluted 10-100 times with sterile water. Counting the disease symptoms of the fritillaria bulb: the surface turns yellow, and the surface has yellow spots and is rotten. Further obtain the control effect of each bacterial liquid on the fritillaria root rot.
The gliocladium roseum antagonizes pathogenic bacteria mainly through the action of heavy parasitism, the culture condition of the substrate has very high organic carbon content, the germination of spores and the growth of hyphae are facilitated, and the gliocladium roseum can be endogenously produced in plants, so the effect in practical application is better.
The soilless substrate used for cultivation in the embodiment is obtained by uniformly mixing turfy soil, rice husk charcoal and vermiculite according to the volume ratio of 3:1: 1.
1. Experiment grouping
Experimental groups: a symbiotic system bacterial liquid consisting of gliocladium roseum and demosaicin;
control group 1: gliocladium roseum strain solution;
positive control group: bacillus amyloliquefaciens liquid;
blank control group: and (5) sterilizing water.
2. Procedure of experiment
(1) Soaking the fritillaria bulb with white, smooth and disease-free surface in 0.2% mercuric chloride, sterilizing the surface for 20 minutes, and cleaning with clear water. Culturing the cleaned Bulbus Fritillariae Cirrhosae seed bulbs in sterilized soilless substrate at room temperature under relative humidity of 60%, and repeating for 3 times each 50 Bulbus Fritillariae Cirrhosae seed bulbs.
(2) Respectively diluting the culture solution obtained from experimental group, control group 1 and positive control group by 100 times, wherein the bacterial count or spore count of the control bacterial solution of experimental group and control group 1 is 104～105The number of bacteria in positive control group control bacteria liquid is 106-107Per ml was poured into the corresponding group; pouring equal volume of sterilized water into the blank control group;
(3) after 5 days, all the fusarium oxysporum bacterial liquid is poured into the liquid in groups (the bacterial count or the spore count is 10)6-107Ml), after 10 days, 1-time pouring fusarium oxysporum bacterial liquid (the number of bacteria or spores is 10)6-107In ml). And (5) counting the disease condition after 30 days, and carrying out statistical analysis by SPSS 22 software.
3. Results of the experiment
Counting the number of the pathogenic shellfish seeds of each group, wherein the result is shown in figure 1, and the incidence rate of the experimental group is (10 +/-3.0)%; the morbidity of the control group 1 is (20 +/-2.6)%, the morbidity of the positive control group is (40 +/-3.5)%, and the morbidity of the blank control group is (90 +/-3.3)%, compared with the blank control group, the morbidity of the experimental group, the control group 1 and the positive control group is obviously reduced, which indicates that the rhizopus rot caused by fusarium pathogenic bacteria can be prevented by the gliocladium roseum and tesserae mosaica symbiotic system bacterial liquid, the gliocladium roseum bacterial liquid and the bacillus amyloliquefaciens bacterial liquid; the incidence of the experimental group was significantly reduced compared to the control group 1 and the positive control group. The results show that the gliocladium roseum and the bacillus amyloliquefaciens can prevent and treat root rot, but the prevention and treatment effect of the fungus gliocladium roseum is stronger than that of the fungus bacillus amyloliquefaciens, the reason that the prevention and treatment effect of the fungus bacillus amyloliquefaciens is weakened can be that the purpose of antagonizing pathogenic bacteria is achieved by the bacillus amyloliquefaciens mainly through secreting bacteriostatic substances, the bacteriostatic substances are diluted in a matrix along with watering, the content of the bacteriostatic substances is low, the bacteriostatic substances are easy to degrade, and the prevention and treatment effect of the bacteriostatic substances is weakened. The control effect of the symbiotic system of the gliocladium roseum and the gliocladium tamariiformis is obviously better than that of the gliocladium roseum and the gliocladium amyloliquefaciens which are independently used, so that the gliocladium roseum and the gliocladium tamariiformis form a good composite microbial agent and can obviously control the occurrence of fritillaria root rot.
Example II prevention and treatment effects of Gliocladium roseum fhnzm and Thymus damascens BMMSJS-1 composite microbial inoculum on Fritillaria kansuensis root rot
In order to further evaluate the prevention and treatment effects of the gliocladium roseum fhnzm and the tesserae mosaic bacterium BMMSJS-1 composite bacterium liquid on the root rot of the fritillaria kansuensis, the field transplanted fritillaria are taken as experimental objects in the embodiment, and the embodiment is divided into three groups which are respectively an experimental group, a blank control group and a traditional method control group. The experimental group is realized by four steps of irrigating the fritillaria seedling culture stage with the compound microbial inoculum bacterial liquid for 3 times and coating the fritillaria small seed stems with the compound microbial inoculum bacterial liquid before transplanting; the fritillaria cultivation method of the blank control group is the same as that of the experimental group, but the compound microbial inoculum liquid is not irrigated; the control group is cultured by a traditional method in the traditional method, and the complex microbial inoculum liquid is not irrigated in the culturing process; the method comprises the following specific operation steps of counting the incidence rate and the green turning rate of the root rot of an experimental group, a blank control group and a traditional method control group:
the main difference between the simulated cultivation test base of the county city horse slope village (with an elevation of 2600 meters) cultivated in the elm base of the present embodiment and the domestication cultivation test base of the city horse gorge town (with an elevation of 1800 meters) cultivated in Huating is that the temperature difference caused by the elevation, the rainfall period and other factors influencing the occurrence of fritillaria diseases are similar. During cultivation, the two bases are respectively provided with 70% of sunshade nets, and the land is a fritillaria bulb continuous cropping land.
The composite microbial inoculum solutions used in the embodiment are all gliocladium roseum fhnzm and demosaicin tima delbrueckii BMMSJS-1 composite microbial inoculum solutions. The number of spores of the composite bacterial liquid is 106-107Per ml, diluted 100 times when used, with spore number of 104-105/ml
The mixed substrate used for cultivation in this example is seedling turfy soil: granular vermiculite: the rice husk charcoal is obtained by mixing the rice husk charcoal in a volume ratio of 3:2: 1. The incidence of diseases is as follows: counting in the middle and last ten days 4 months after seedling emergence;
green turning rate: in the middle and last ten days of the next 4 months (the incidence rate is counted in the next year), as the rainy season in Gansu is mainly concentrated in 7-9 months, although the fritillaria is fallen to seedlings, the fritillaria buried in the soil is easy to suffer from root rot, the green return rate in the next year is reduced, and the green return rate in the next year is counted.
Experimental groups:
1. seed of Fritillaria przewalskii Mak in Gansu province breaks dormancy
Collecting seeds of fritillaria kansuensis when the half-yellow fruits are not completely mature and cracked in 6 months in the 1 st year, carrying out sand storage at the temperature of 16 ℃ for 14h/8 ℃ for 10h for 3 months at variable temperature until the embryo yield reaches more than 0.6, and then carrying out sand storage at the temperature of 2 ℃ for 2 months at constant temperature until the embryo yield reaches more than 0.8.
2. Seed sowing
Sterilizing the seeds after post-ripening for 10min with 0.2% mercuric chloride, washing with sterile water, sowing in sterilized mixed matrix, culturing in greenhouse at 1 month in 2 years, and controlling the minimum temperature of the greenhouse to be more than 0 deg.C.
3. Inoculating, culturing, and storing at low temperature
The seed begins to emerge about 20 days after sowing, the composite microbial inoculum is irrigated for 1 time after emergence of seedlings, and the composite microbial inoculum is diluted by 100 times during irrigation until the number of spores is 104-105And/ml, pouring seedlings in 5 months in 2 years, wherein the size of the bulblet is 2-3 mm. And (3) storing the bulbils irrigated with the composite microbial inoculum liquid at the low temperature of 2 ℃ for 3 months.
4. Sowing the seeds to the small seed stems for harvesting
Sowing the bulblets after low-temperature storage into the same mixed matrix after sunstroke at 8 months in the 2 nd year, culturing under greenhouse condition, and controlling the highest temperature of the greenhouse<30 ℃ and about 20 days or soSeedling emergence, irrigating the composite microbial inoculum after seedling emergence, diluting by 100 times during irrigation until the number of spores is 104-105And/ml, growing for 12 months, and gradually reducing the temperature in the greenhouse until the fritillaria is transplanted. At the moment, the bulblet is 3-4 mm in size and can be transplanted as a microspecite in a field.
5. Low-temperature storage and coating of small seed stems
And (4) burying the small seed stems obtained in the step (4) in the mixed matrix irrigated with the composite microbial inoculum solution, and storing at a low temperature of 2 ℃ for 3 months. Taking out in 3 months of 3 years, soaking in a compound bacterial liquid containing 0.05% hydroxypropyl methyl cellulose for coating, soaking for 20min, taking out, drying in the shade, and transplanting in the field.
6. Transplanting in field
And (4) transplanting the small seed stems obtained in the step (5) to a simulated cultivation test base of the city Mapo county in Ulcenter and a domestication cultivation test base of the city Maxia towns in Huating. Transplanting 5000 seedlings in 3 rd year (2018) in the first 4 th month (pseudo-habitat cultivation base) and the last 3 th month (domesticated cultivation base), wherein the transplanting row spacing is 10cm, and the plant spacing is 5 cm. And after seedling emergence, counting the incidence rate of the root rot in the last 4-5 months in 2018, and counting the green-turning rate in the last 4-5 months in 2019.
Blank control group:
except that the cultivation process of the fritillaria seeds is not carried out by irrigating the compound microbial inoculum (4 times of cultivation), all cultivation methods of the control group 1 are the same as those of the experimental group, the growth condition of the fritillaria is observed, the incidence rate of root rot is counted in the last 4-5 months in 2018, and the green return rate is counted in the last 4-5 months in 2019.
Control group of conventional method:
adopting a field direct seeding method to raise seedlings (different from methods of an experimental group and a blank control group), collecting seeds of the fritillaria kansuensis in 1 year (2015 year) and 6 months when the half yellow fruits are not completely mature and cracked, selecting a slope field to raise ridges and sow the collected seeds, covering straws, pine needles and the like for preserving soil moisture, and seedling emergence at the bottom of 3 months in 2 years (2016 years) and seedling inversion at the bottom of 6 months; in the 3 rd year (2017), 3 month end seedlings emerge, 6 month end seedlings fall, 10 months are taken out, 2 years of small stems of the fritillaria are harvested, the small stems are respectively transplanted to a city MaoPo country simulation cultivation test base in Yuzhong county and a city Maxia town domestication cultivation test base in Huating, 70% of sunshade nets are respectively erected on the two bases, the field is a fritillaria heavily-stubbled land, the transplanting row spacing is 10cm, the planting distance is 5cm, 3 last ten days to 4 last ten days of 3 months in the 4 th year (2018) after transplantation, the incidence rate of root rot is counted in 4 to 5 last ten days, and the green return rate is counted in 4 to 5 last ten days of 2019.
TABLE 1 statistics of the growth of fritillaria under the same cultivation method
| Growth conditions | Experimental group | Blank control group |
| End of first life | 2-3 mm in diameter | Diameter of 1-1.5 mm |
| End of second growth phase | The diameter is 3-4 mm | The diameter is 2-2.5 mm |
| Emergence rate in the second growth period (%) | 70±6.3* | 20±3.2 |
Note: means significant difference in comparison between experimental and control groups
TABLE 2 growth of fritillaria with different cultivation methods and cultivation bases
Note:#shows that the comparison between different cultivation bases has significant difference
Means significant difference in comparison between experimental and control groups
The growth conditions of the fritillaria under the same cultivation method are shown in table 1, and the fritillaria seeds in the experimental group are sown in the mixed matrix, so that the growth condition is good, and no diseases occur; compared with the experimental group, the seedling diameter and the emergence rate of the blank control group are lower than those of the experimental group, so that the composite microbial inoculum provides nitrogen nutrition for the seedling of the fritillaria kansuensis, the growth is promoted, and sufficient nutrition provides guarantee for the development of the heart bud.
The growth conditions of the fritillaria in different cultivation methods and cultivation bases are shown in table 2, and the fritillaria can be obtained from the table 2, so that compared with a control group in the traditional method, the incidence of diseases in an experimental group is lower, and the green turning rate is higher; compared with the Huating pavilion domestication cultivation base, the planting base in the elm is lower in incidence rate and higher in green turning rate.
In conclusion, compared with the fritillaria seedlings without the composite microbial inoculum, the fritillaria seedlings with the composite microbial inoculum liquid has the advantages that the incidence rate of the root rot is remarkably reduced, the green turning rate is remarkably improved, and the composite microbial inoculum liquid has a protection effect on the root of fritillaria and can prevent the occurrence of the root rot of fritillaria.
Example III application of Gliocladium roseum fhnzm and Thymus amabilis BMMSJS-1 composite bacterial agent to prevention and treatment of angelica root rot and pockmark disease
In order to evaluate the prevention and treatment effects of the compound microbial inoculum on the root rot and the pockmark disease of the angelica, the embodiment is divided into two groups, namely an experimental group and a control group. The experimental group is realized by three steps of irrigating the Chinese angelica seedling culture stage with the compound microbial inoculum bacterial liquid for 1 time, coating the seedlings with the compound microbial inoculum bacterial liquid before transplanting, and irrigating roots of the seedlings with the compound microbial inoculum bacterial liquid for 1 time after the seedlings are transplanted in a field; the control group is not irrigated with the compound microbial inoculum solution;
note: because the root rot of angelica is concentrated in early spring seasons and the environmental conditions of dry cooling and wet cooling are easy to cause diseases, the disease occurrence condition of the root rot is counted in the middle of 6 months; the occurrence and the disease degree of the leprosy can not cause plant death, so the statistics of the disease degree of the leprosy is not carried out in the experiment, and only the disease condition is counted.
The mixed substrate used for cultivation in this example is seedling turfy soil: granular vermiculite: the rice husk carbon is obtained by mixing the rice husk carbon with the volume ratio of 3:2: 1.
The number of fungal spores of the composite bacterial liquid irrigated by the embodiment is 106-107The volume of the solution is 10 times that of the solution when diluted and is between 10 times that of the solution after dilution5-106/ml。
Experimental groups:
1. seed sowing
Harvesting angelica sinensis seeds from the bottom of 10 months to the beginning of 11 months in the 1 st year, sterilizing for 10min by 0.2 percent of mercury bichloride, washing with sterile water, sowing in a sterilized mixed matrix in 12 months, culturing under a greenhouse condition, controlling the lowest temperature of the greenhouse to be more than 0 ℃, starting seedling emergence in about 10 days, irrigating gliocladium roseum and tesseradilla moellendorfii composite bacterial liquid for 1 time after seedling emergence, controlling the seedling stage to be 3-4 months according to the seedling growth, and gradually reducing the temperature in the middle and last ten days of 3 months in the 2 nd year to enable the leaves to yellow, and starting and transplanting.
2. Seedling coating field planting
And (3) lifting seedlings at the end of 3 months in the 2 nd year, soaking the seedlings in a gliocladium roseum and sambucus sumatra composite microbial inoculum containing 0.05 percent of hydroxypropyl methyl cellulose for coating, taking out the seedlings after soaking for 20min, drying the surface water in the shade, transplanting the seedlings in a field, experimentally locating the seedlings in Ten Town (the altitude of 2200 m) of Min county, transplanting the seedlings in a field block with the Chinese angelica for multiple crops, transplanting the seedlings at the row spacing of 20cm and the plant spacing of 10cm, and carrying out single-seedling hole planting on the seedlings to obtain 5000 seedlings.
3. Irrigating roots after seedling emergence
Sprouting begins about 10 days after the coated seedlings are planted, when the seedlings grow to 3-4 leaves in the early 5 months of the 2 nd year, the broomcorn gloriopsis rosea and rimonapris timonii compound bacterium liquid is diluted by 10 times and the number of the fungus spores is 10 after dilution5-106/ml。
4. Disease statistics
In 6 th month in 2 nd year, counting the occurrence of root rot, picking radix Angelicae sinensis in 11 th month in 2 nd year, counting the occurrence of mottle, and counting data without bolting radix Angelicae sinensis.
Control group:
and (3) in the middle ten days of 6 months, directly seeding and raising seedlings in the field, wherein the seedling period is controlled to be about 3 and a half months, the seedlings are lifted in 10 months, bundling and stacking are carried out, transplanting is carried out at the bottom of 3 months in the next year, and the transplanting place and the transplanting method are the same as those in the embodiment 3. In late 6 months of the next year, counting the incidence of root rot, digging angelica sinensis at the beginning of 11 months of the next year, counting the incidence of rough mouth disease and root rot, and counting data without bolting the angelica sinensis.
TABLE 3 incidence of root rot and Kaschin-Beck disease in Angelica sinensis treated differently
| Incidence of disease | Experimental group | Control group |
| Root rot (%) | 0* | 30±4.3 |
| Mottle disease (%) | 8±1.8* | 20±3.6 |
Note: means significant difference in comparison between experimental and control groups
The occurrence of root rot and pockmark disease of angelica treated differently is shown in table 3, and it can be seen from table 3 that the incidence of root rot and pockmark disease of angelica with the compound bacterial liquid irrigated in the experimental group is significantly lower than that of root rot and pockmark disease of angelica with the compound bacterial liquid not irrigated in the control group. Therefore, the compound microbial inoculum can be used for preventing and treating the root rot and the pockmark disease of the angelica.
In conclusion, the composite microbial inoculum bacterial liquid claimed by the invention can fix nitrogen, promote the growth of seedlings, preferentially symbiote with the seedlings in a sterile soilless substrate seedling raising mode under facility conditions, and breed high-quality seedlings carrying probiotics. In the field cultivation stage, the spread way of pathogenic fungi can be cut off by means of seedling coating and root irrigation after cultivation, the pathogenic fungi can be inhibited and killed, and the nematodes and underground pests can be inhibited and killed. The fungus gliocladium roseum in the compound microbial inoculum liquid has the functions of inhibiting and killing bacteria, fungi, nematodes and underground pests, has the main action mechanisms of heavy parasitism and secretion of antagonistic substances, can be propagated in an application environment through spores, and has better effect on the growth promotion of alpine shade plants and the prevention of plant diseases and insect pests than probiotic bacteria. The composite microbial inoculum bacterial liquid can be used for preventing and treating diseases such as root rot and pockmark disease of the alpine shade plants and promoting the growth of the alpine nitrogen-rich plants.
Claims (9)
1. A composite microbial preparation comprises Gliocladium roseum and mosaic bacterium delmophila.
2. The complex microbial inoculant according to claim 1, wherein the Gliocladium roseum is Gliocladium roseum (Clinostachysossea) fhnzm, deposited at the China general microbiological culture Collection center with the collection number of CGMCC No. 17070.
3. The complex microbial inoculant according to claim 1 or 2, wherein the Marasmius timonii is Marasmius timonii (Massiitiarimonae) BMMSJS-1, deposited at the China general microbiological culture Collection center with the collection number of CGMCC No. 18668.
4. The use of the complex microbial inoculum of claim 3 in inhibiting fusarium pathogens.
5. The use according to claim 4, wherein the Fusarium pathogenic bacteria is Fusarium solani or Fusarium oxysporum.
6. The application of the composite bacterial agent of claim 3 in preventing and treating plant diseases caused by fusarium pathogenic bacteria.
7. The use according to claim 6, wherein the plant is a medicinal plant, and the disease is one or more of rot, blight and numb mouth disease caused by fusarium pathogenic bacteria.
8. A pesticide for controlling plant diseases caused by fusarium pathogenic bacteria, which is characterized by comprising a composite microbial inoculum consisting of the gliocladium roseum and the pimavalia rimonansis according to claim 3.
9. The pesticide of claim 8, wherein the gliocladium roseum and gliocladium intramurae complex inoculant has a gliocladium roseum spore number of 106～107/ml。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010336121.9A CN111471598B (en) | 2020-04-25 | 2020-04-25 | Gliocladium roseum and pimavalia rimonaris composite microbial agent and application thereof in prevention and control of diseases and pests |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010336121.9A CN111471598B (en) | 2020-04-25 | 2020-04-25 | Gliocladium roseum and pimavalia rimonaris composite microbial agent and application thereof in prevention and control of diseases and pests |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111471598A true CN111471598A (en) | 2020-07-31 |
| CN111471598B CN111471598B (en) | 2021-02-19 |
Family
ID=71755631
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010336121.9A Active CN111471598B (en) | 2020-04-25 | 2020-04-25 | Gliocladium roseum and pimavalia rimonaris composite microbial agent and application thereof in prevention and control of diseases and pests |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111471598B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111919692A (en) * | 2020-08-05 | 2020-11-13 | 苏宗然 | Planting method for green prevention, control and efficient production of chicory for northern powder |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102174460A (en) * | 2011-01-23 | 2011-09-07 | 青岛科技大学 | Gliocladium roseum chlamydospore and method for producing wettable powder thereof |
| WO2014201044A2 (en) * | 2013-06-10 | 2014-12-18 | The Regents Of The University Of California | Plant growth-promoting microorganisms and methods of use thereof |
| CN106244499A (en) * | 2016-09-20 | 2016-12-21 | 中节能六合天融环保科技有限公司 | One strain Di Monei Marseille bacterium new strains and application thereof |
| CN106305792A (en) * | 2016-05-19 | 2017-01-11 | 北京启高生物科技有限公司 | Clonostachys rosea-bacillus subtilis composite wettable powder |
| EP3254565A1 (en) * | 2016-06-06 | 2017-12-13 | Etablissements J. Soufflet | Microbial strains for biologically controlling fusarium head blight |
| CN109762743A (en) * | 2019-01-30 | 2019-05-17 | 河南省农业科学院植物保护研究所 | Gliocladium roseum, its solid fermentation microbial inoculum and its application |
| CN110172408A (en) * | 2019-04-10 | 2019-08-27 | 甘肃省科学院生物研究所 | The endogenetic fungus of one plant of Chinese podophyllum root and its application |
| EP3626815A1 (en) * | 2017-05-18 | 2020-03-25 | Obshchestvo S Ogranichennoy Otvetsvennostyu "Ekogen" | Strain of microorganism clonostachys rosea f. catenulata as a biofungicide, plant growth stimulant and metabolite producer for agricultural use |
-
2020
- 2020-04-25 CN CN202010336121.9A patent/CN111471598B/en active Active
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102174460A (en) * | 2011-01-23 | 2011-09-07 | 青岛科技大学 | Gliocladium roseum chlamydospore and method for producing wettable powder thereof |
| WO2014201044A2 (en) * | 2013-06-10 | 2014-12-18 | The Regents Of The University Of California | Plant growth-promoting microorganisms and methods of use thereof |
| CN106305792A (en) * | 2016-05-19 | 2017-01-11 | 北京启高生物科技有限公司 | Clonostachys rosea-bacillus subtilis composite wettable powder |
| EP3254565A1 (en) * | 2016-06-06 | 2017-12-13 | Etablissements J. Soufflet | Microbial strains for biologically controlling fusarium head blight |
| CN106244499A (en) * | 2016-09-20 | 2016-12-21 | 中节能六合天融环保科技有限公司 | One strain Di Monei Marseille bacterium new strains and application thereof |
| EP3626815A1 (en) * | 2017-05-18 | 2020-03-25 | Obshchestvo S Ogranichennoy Otvetsvennostyu "Ekogen" | Strain of microorganism clonostachys rosea f. catenulata as a biofungicide, plant growth stimulant and metabolite producer for agricultural use |
| CN109762743A (en) * | 2019-01-30 | 2019-05-17 | 河南省农业科学院植物保护研究所 | Gliocladium roseum, its solid fermentation microbial inoculum and its application |
| CN110172408A (en) * | 2019-04-10 | 2019-08-27 | 甘肃省科学院生物研究所 | The endogenetic fungus of one plant of Chinese podophyllum root and its application |
Non-Patent Citations (4)
| Title |
|---|
| ZERIHUN A.DEMISSIE ET AL.: ""Profiling of the Transcriptomic Responses of Clonostachys rosea Upon Treatment With Fusarium graminearum Secretome"", 《FRONTIERS IN MICROBIOLOGY》 * |
| 杨恩东等: ""马赛菌属细菌研究进展"", 《微生物学通报》 * |
| 杨蕊等: ""粉红粘帚霉对玉米茎基腐病的抑菌防病作用"", 《河南科技学院学报》 * |
| 高航: ""粉红粘帚霉菌对四种林果枝干病原菌的生防作用研究"", 《中国优秀硕士学位论文全文数据库 农业科技辑》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111919692A (en) * | 2020-08-05 | 2020-11-13 | 苏宗然 | Planting method for green prevention, control and efficient production of chicory for northern powder |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111471598B (en) | 2021-02-19 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109971680B (en) | Preparation and application of compound microbial agent with disease-resistant and growth-promoting functions | |
| CN103271095B (en) | Composite microbial agent for preventing and treating fruit tree replant disease and disease prevention method thereof | |
| CN100418400C (en) | Standard planting method of medicinal lycoris radiata | |
| US7405181B2 (en) | Bio-fertilizer composition for promoting growth or orchid plants and application | |
| CN111418418A (en) | Green and safe pest control method | |
| CN113564054B (en) | Method for improving plant disease resistance by using beauveria bassiana blastospore | |
| CN104686196B (en) | Method for preserving and separating toadstool strain through sporocarp dried in shade | |
| CN111602567A (en) | Compound grass planting management method for fallen leaf orchard | |
| JPH0819407B2 (en) | Soil conditioner for plant cultivation, production method and use method thereof | |
| Tu | The role of white mold‐infected white bean (Phaseolus vulgaris L.) seeds in the dissemination of Sclerotinia sclerotiorum (Lib.) de Bary | |
| CN111471598B (en) | Gliocladium roseum and pimavalia rimonaris composite microbial agent and application thereof in prevention and control of diseases and pests | |
| CN110129242B (en) | Continuous cropping resistant composite microbial preparation and preparation method thereof | |
| CN116790417A (en) | Pseudomonas capable of inhibiting pathogenic bacteria of various plants and having growth promoting effect on various crops, screening method and application | |
| CN112522110B (en) | Microbial agent suitable for dendrobium wilt and application thereof | |
| JP2022077963A (en) | Plant seedling, seedling cultivation method, culture soil, and method of growing plant | |
| CN109161486B (en) | Preparation method of biocontrol trichoderma strain inoculant and application of biocontrol trichoderma strain inoculant in tobacco planting | |
| CN111492893A (en) | Method for cultivating agaricus bisporus with high content of trace elements such as organic selenium, zinc, calcium and magnesium | |
| CN111512912A (en) | Cultivation method for reducing incidence of bacterial leaf blight of rice | |
| CN104087542A (en) | Biocontrol strain for preventing and controlling fusarium wilt of watermelon and application thereof | |
| CN110024611B (en) | Method for improving disease resistance of betel nuts through arecoline treatment and application | |
| JP7038451B1 (en) | Plant seedlings, seedling raising methods, hilling, and plant growing methods | |
| CN111248047B (en) | Method for planting arborvitae artificial forest by using cork oak | |
| CN106386096A (en) | Method for planting wild affine cudweed | |
| Rai et al. | Potential Use Of Animal Manures In Managing Phytophthora Wilt Of Chilli Caused By Phytophthora Capsici | |
| CN116622542A (en) | Bacillus tropicalis and application thereof in ginger |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| CB03 | Change of inventor or designer information | ||
| CB03 | Change of inventor or designer information |
Inventor after: Yang Tao Inventor after: Wang Zhiye Inventor after: Ma Yonggui Inventor after: Yang Hui Inventor after: Xie Huichun Inventor after: Zhao Jiang Inventor after: Ma Yulan Inventor after: Fang Yanhao Inventor after: Li Xin Inventor before: Yang Tao Inventor before: Wang Zhiye Inventor before: Yang Hui Inventor before: Zhao Jiang Inventor before: Fang Yanhao Inventor before: Li Xin |
|
| TA01 | Transfer of patent application right | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20210128 Address after: 730000 No. 197 South Dingxi Road, Chengguan District, Gansu, Lanzhou Applicant after: Institute of Biology, Gansu Academy of Sciences Applicant after: QINGHAI NORMAL University Address before: 730000 No. 197 South Dingxi Road, Chengguan District, Gansu, Lanzhou Applicant before: Institute of Biology, Gansu Academy of Sciences |
|
| GR01 | Patent grant | ||
| GR01 | Patent grant |