CN105779305B - Special compound microbial agent for eliminating successive cropping obstacles of radix pseudostellariae and preparation method thereof - Google Patents
Special compound microbial agent for eliminating successive cropping obstacles of radix pseudostellariae and preparation method thereof Download PDFInfo
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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- C12N1/20—Bacteria; Culture media therefor
Abstract
The invention discloses a special compound microbial agent for eliminating successive cropping obstacles of radix pseudostellariae, which consists of general beneficial bacteria and indigenous beneficial bacteria in a volume ratio of 4: 6; the general beneficial bacteria are prepared by mixing lactic acid bacteria, photosynthetic bacteria, saccharomycetes and actinomycetes according to the volume ratio of 1:1:1: 1; the indigenous beneficial bacteria are prepared by mixing Burkholderia, pseudomonas and bacillus according to the volume ratio of 4:1: 1. The invention also discloses a preparation method of the special compound microbial agent for eliminating the continuous cropping obstacle of the radix pseudostellariae, which comprises the following steps: step one, screening the indigenous antagonistic bacteria; step two, preparing general composite bacterial liquid; step three, activating and expanding culture of indigenous beneficial bacteria; step four, preparing the compound microbial agent. The special compound microbial agent prepared by the invention has good rhizosphere colonization capability, is safe and reliable, has obvious prevention and treatment effects on continuous cropping obstacles and soil-borne diseases of the radix pseudostellariae, and has no pathogenic infection capability on other crops in the succeeding crop.
Description
Technical Field
The invention relates to the technical field of microorganism and biological control, in particular to a special compound microorganism bacterium agent for eliminating successive cropping obstacles of radix pseudostellariae and a preparation method thereof.
Background
The continuous cropping obstacle refers to the phenomenon that the same plants are planted in the same land for many years continuously even under normal field management measures, so that the growth and development of the plants are poor, the plant diseases and insect pests are serious, and the yield and the quality are reduced, and is also called a chemical infection and self-toxicity phenomenon or a replanting disease. The medicinal plants are generally prohibited from continuous cropping, and according to statistics, about 70 percent of root tuber medicinal materials have continuous cropping obstacles in different degrees, which seriously hinders the sustainable development of the modern pharmaceutical industry in China.
Years of field observation shows that the continuous cropping radix pseudostellariae plants are poor in growth and development, underground root tubers cannot normally expand, and the seeds can be replanted in the same field at intervals of 2-4 years after being harvested. Earlier researches have considered that the unbalanced rhizosphere microbial community structure under continuous cropping, namely the reduction of beneficial bacteria and the mass propagation and growth of pathogenic bacteria, are main factors causing the continuous cropping obstacle of the radix pseudostellariae. The successive cropping of the radix pseudostellariae also causes the problems of soil acidification, self-toxic substance release, soil adhesion and hardening, water and fertilizer retention capacity reduction and the like, and meanwhile, under the condition that the mechanism for forming successive cropping obstacles of the radix pseudostellariae is not clear, farmers try to maintain the yield by abusing pesticides and fertilizers, the effect is often poor, and the problems of increased production cost, serious fertilizer loss, pesticide residues and the like are caused. Therefore, the important significance is brought to the exploration of an environment-friendly continuous cropping obstacle reduction measure.
The method is used for improving deteriorated continuous cropping soil through a microbial agent or a biological organic fertilizer, improving the diversity level of soil microorganisms, and repairing and reconstructing a healthy soil ecosystem and is considered to be a main way for solving the continuous cropping obstacle of plants. As shown in figure 1, the inventor selects a plurality of highly pathogenic bacteria such as fusarium oxysporum, fusarium moniliforme, fusarium anklebone and the like from diseased radix pseudostellariae plants and continuous cropping soil in an earlier stage, and the plurality of pathogenic bacteria show host specificity, namely only infect the radix pseudostellariae and do not infect other crops such as corn, soybean, rice and the like in later stages. Therefore, beneficial bacteria capable of antagonizing the special pathogenic bacteria of the radix pseudostellariae can be separated and screened to effectively prevent and control the replantation disease of the radix pseudostellariae. Earlier researches also find that commercial microbial agents or bacterial fertilizers sold in the market have very limited effect on preventing and treating the continuous cropping obstacle of the radix pseudostellariae, which may be related to rhizosphere colonization capacity of beneficial bacteria, and the beneficial bacteria are not suitable for growing in the continuous cropping soil of the radix pseudostellariae. The continuous cropping of the radix pseudostellariae causes obvious deterioration of the soil environment, and the radix pseudostellariae continuously releases various secondary metabolites and drug effect components which can cause that beneficial bacteria in a commercial microbial agent cannot colonize in the rhizosphere of the radix pseudostellariae, so that the host specificity of the soil-borne pathogenic bacteria of the radix pseudostellariae and the particularity of the continuous cropping soil environment need to be comprehensively considered when the microbial agent is adopted to prevent and treat the replantation diseases of the radix pseudostellariae.
In view of the above, the present inventors have researched and designed a special composite microbial agent for eliminating the continuous cropping obstacle of radix pseudostellariae and a preparation method thereof, and have generated the present invention.
Disclosure of Invention
The invention aims to provide a special compound microbial agent for eliminating successive cropping obstacles of radix pseudostellariae, which is used for biologically preventing and treating the successive cropping obstacles of the medicinal plant radix pseudostellariae and has double purposes of promoting root tuber growth and preventing and treating diseases.
The invention also aims to provide a preparation method of the special compound microbial agent for eliminating the continuous cropping obstacle of the radix pseudostellariae, which is prepared by mixing universal beneficial bacteria and indigenous beneficial bacteria in an optimized proportion, and aims to ensure the successful colonization of the beneficial bacteria at the rhizosphere of the radix pseudostellariae, activate soil nutrients, degrade autotoxic substances released by the radix pseudostellariae and prevent and treat specialized pathogenic bacteria of the radix pseudostellariae.
In order to achieve the purpose, the technical scheme adopted by the invention for solving the technical problem is as follows:
a special compound microbial agent for eliminating successive cropping obstacles of radix Pseudostellariae comprises general beneficial bacteria and indigenous beneficial bacteria at a volume ratio of 4: 6; the general beneficial bacteria are prepared by mixing lactic acid bacteria, photosynthetic bacteria, saccharomycetes and actinomycetes according to the volume ratio of 1:1:1: 1; the indigenous beneficial bacteria are prepared by mixing Burkholderia, pseudomonas and bacillus according to the volume ratio of 4:1: 1.
In a preferred embodiment, the lactic acid bacteria are one or more of lactobacillus plantarum, streptococcus lactis and lactobacillus casei.
In a preferred embodiment, the photosynthetic bacteria are one or both of rhodopseudomonas palustris and rhodobacter sphaeroides.
In a preferred embodiment, the yeast is one or two of saccharomyces cerevisiae and candida utilis.
In a preferred embodiment, the actinomycetes is one or both of Streptomyces griseus and Streptomyces albus.
As a preferred embodiment, the Burkholderia plantarii is one or both of Burkholderia plantarii (Burkholderia plantarii sp. nov., at least one of the Burkholderia cepacia complex including biocontrol and cystic fibrosis-related isolates. Tom Coense, Ehwarhira mahogany, Deborah Henry, et. International Journal of Systematic and evolution Microbiology, 2001(51):1481-1490.) and New Burkholderia cepacia.
As a preferred mode of embodiment, the Pseudomonas is one or more of Pseudomonas fluorescens, Pseudomonas putida, Pseudomonas proteins (Pseudomonas proteins sp. nov., wild plant-detecting bacterium reducing the biocontrol compounds 2, 4-diacetylphospherol and pyro lysis. Alban Rameter, Michel Frapol, Marion Fischer-Le Saux. et. systematic and Applied Microbiology, 2011(34): 180-.
As a preferable mode of embodiment, the bacillus is one or more of bacillus amyloliquefaciens, bacillus subtilis, bacillus cereus, bacillus pumilus, bacillus megaterium and bacillus mycoides.
A preparation method of a special compound microbial agent for eliminating successive cropping obstacles of radix pseudostellariae comprises the following steps:
step one, screening the indigenous antagonistic bacteria:
separating pseudostellaria root pseudomonas and bacillus by adopting a pseudomonas selective culture medium and a bacillus selective culture medium respectively, and separating autotoxic substance degrading bacteria by adopting an inorganic culture medium;
the formula of the pseudomonas selective culture medium is as follows: weighing 20g of soybean-casein digest agar culture medium, adding 495.5mL of double distilled water, heating and stirring, fully dissolving, then adding 1mL of 0.1% (wt/vol) crystal violet stock solution, carrying out autoclaving at 121 ℃ for 15min, cooling to about 50 ℃, adding 3.5mL of 5% (wt/vol) furantoin stock solution into the culture medium, using N, N-dimethylformamide as a solvent for the furantoin stock solution, and carrying out back plate cooling for later use;
the bacillus selective culture medium comprises the following components in percentage by weight: 5.0g peptone, 1.5g yeast extract, 1.5g beef extract, 5.0g NaCl, 20g agar per liter, pH 7.2;
the formula of the inorganic culture medium is as follows: NH per liter4NO3 1.0g，NaCl 0.5g，K2HPO4·3H2O 1.5g，MgSO4·7H2O 0.5g，(NH4)2SO40.5g, pH 7.2; weighing each component, dissolving the components in 1000mL of distilled water, sterilizing the components for 15min at 121 ℃ under high pressure, cooling the components to 45-50 ℃, adding autotoxic substances identified in the radix pseudostellariae continuous cropping soil, wherein the autotoxic substances comprise gallic acid, coumaric acid, 3, 4-dihydroxybenzoic acid, p-hydroxybenzoic acid, vanillic acid, syringic acid, vanillin, ferulic acid and benzoic acid, the total concentration of the mixed phenolic acid is 1000mg/L, inoculating 10% radix pseudostellariae rhizosphere soil suspension, enriching and culturing for several generations, and screening beneficial bacteria with autotoxic substance degradation capability and antagonistic effect on pathogenic bacteria of the radix pseudostellariae;
step two, preparing general composite bacterial liquid:
adding 300g of brown sugar and 50mL of MS mother liquor into 18L of well water or tap water which is placed for at least 2 days, adding 1L of activated EM liquid strain, placing the well water or the tap water in a room at 35-37 ℃ for closed fermentation for 2-3 weeks, opening a closed container opening every other day from the second day of fermentation to deflate and decompress once, and opening the container opening to smell sour and sweet taste, which is more and more obvious and indicates that fermentation expanding culture is successful, so as to obtain EM composite bacterial liquid for later use; the successful finished product fermentation liquor has acid and sweet taste, the pH value is 3.0-4.0, and the viable count is more than 100 hundred million/mL;
step three, indigenous beneficial bacteria activation and enlarged culture:
activating strains: respectively inoculating the screened Pseudomonas fluorescens, Pseudomonas putida, Pseudomonas protegens and Pseudomonas korea into a Pseudomonas selective culture medium, and activating in an incubator at 28-32 ℃ for 12-16 h; respectively inoculating the screened burkholderia and bacillus into LB solid culture, and putting the obtained product into an incubator at 37 ℃ for activation for 8-16 h;
and (3) amplification culture: inoculating activated various indigenous beneficial bacteria into liquid culture for fermentation culture, performing amplification culture on pseudomonas at 28-32 ℃, performing amplification culture on Burkholderia and bacillus at 37 ℃, and performing amplification culture when the viable count is higher than 1.0 × 109Stopping the amplification culture at CFU/mL for later use;
step four, preparing the compound microbial agent:
mixing indigenous beneficial bacteria, namely Burkholderia, pseudomonas and bacillus according to the volume ratio of 4:1:1 to prepare an indigenous beneficial bacteria compound bacterial liquid, namely an IM compound bacterial liquid; and mixing the EM composite bacterial liquid and the IM composite bacterial liquid according to the volume ratio of 4:6 to prepare the special composite microbial agent for the radix pseudostellariae.
The special compound microbial agent has the following advantages:
1. the special compound microbial agent contains general beneficial bacteria and rhizosphere antagonistic bacteria of the radix pseudostellariae, wherein the rhizosphere antagonistic bacteria comprise Burkholderia, pseudomonas and bacillus, which are obtained by screening from rhizosphere soil of the radix pseudostellariae, and the rhizosphere antagonistic bacteria have good rhizosphere colonization capability, are safe and reliable when applied in fields, and have no pathogenic infection capability on other crops (such as corn, soybean, rice, vegetables and the like) in the next period;
2. the indigenous antagonistic bacteria are obtained by a large amount of screening work, are obtained by screening 3000 soil bacteria, and have strong antagonistic effect on special pathogenic bacteria of radix pseudostellariae (such as fusarium oxysporum, fusarium moniliforme, talaromyces tarda and the like).
3. When the indigenous antagonistic bacteria are screened, autotoxic substances with higher concentration are added into a culture medium as selection pressure, and most of screened indigenous beneficial bacteria have degradation and conversion functions on autotoxic substances and drug effect components (such as the pseudostellaria cyclic peptide B) secreted by the root system of the pseudostellaria heterophylla.
4. The indigenous antagonistic bacterium provided by the invention has the phosphate solubilizing capability besides inhibiting pathogenic bacteria, can effectively activate soil nutrients, improves the physicochemical properties of soil, and plays an important role in promoting the absorption of plant nutrients.
5. The invention is verified on the spot for a long time, has reasonable formula, stable effect, obvious effect of preventing and treating the successive cropping obstacle of the radix pseudostellariae and improvement of the yield and the quality of the successive cropping radix pseudostellariae.
Drawings
FIG. 1 shows the colony morphology of pathogenic bacteria isolated from roots of a disease of a plant of radix Pseudostellariae; wherein, A: fusarium oxysporum; b: fusarium moniliforme; c: talaromyces;
FIG. 2 is a photograph of the confrontation culture of the indigenous antagonistic bacteria and the special pathogenic fungi of radix Pseudostellariae (Fusarium moniliforme); wherein, A: only inoculating fusarium moniliforme of pathogenic bacteria; b: indigenous beneficial bacteria are cultured in a confronting manner with the fusarium moniliforme;
FIG. 3 is the determination of the phosphate-solubilizing ability of indigenous beneficial bacteria; wherein, the arrows indicate indigenous beneficial bacteria with phosphate-solubilizing ability, and a transparent ring is formed around the colony.
Detailed Description
The invention is further described with reference to the following drawings and detailed description,
burkholderia cepacia, Pseudomonas koreana and Burkholderia ambifaria have been registered and preserved in the China general microbiological culture Collection center at 28 th 2016 (month 01), with the preservation address of No. 3 Hospital No.1 Homew of North Kyowa, sunward, and the preservation numbers of CGMCC No.12119, CGMCC No.12122, CGMCC No.12121 and CGMCC No.12120, respectively.
Example 1 screening and identification of indigenous antagonistic bacteria
The pseudostellaria root pseudomonas and bacillus are separated by adopting a pseudomonad selective culture medium and a bacillus selective culture medium respectively, and the autotoxic substance degrading bacteria are separated by adopting an inorganic culture medium (containing higher-concentration autotoxic substances of the pseudostellaria).
The antagonistic effect of the randomly selected single colonies on the different culture media on the special pathogenic bacteria (such as fusarium oxysporum, fusarium moniliforme, talaromyces ankara and the like) of the radix pseudostellariae is respectively evaluated by adopting a PDA plate antagonistic method. The bacteria with strong antagonistic ability are subjected to molecular identification by adopting 16s rRNA gene or 16s-23s rRNA interval PCR identification. The 16s rRNA gene PCR amplification upstream and downstream primers are respectively as follows: 27f (5'-AGAGTTTGATCCTGGCTCAG-3') and 1522r (5'-AAGGAGGTGATCCAGCCGCA-3'); the 16s-23s rRNA gene PCR amplification upstream and downstream primers are respectively as follows: 1407f (5'-TTGTACACACCGCCCGTC-3') and 456r (5'-CCTTTCCCTCACGGTACTG-3').
The formula of the culture medium is as follows:
(1) pseudomonas selective culture medium: weighing 20g of soybean-casein digest agar medium, adding 495.5mL of double distilled water, heating and stirring to fully dissolve, then adding 1mL of 0.1% (wt/vol) crystal violet stock solution, carrying out autoclaving at 121 ℃ for 15min, cooling to about 50 ℃, adding 3.5mL of 5% (wt/vol) furantoin stock solution (N, N-dimethylformamide is used as a solvent) into the medium, and carrying out plate inversion cooling for later use.
(2) Bacillus selective culture medium: each liter contained 5.0g peptone, 1.5g yeast extract, 1.5g beef extract, 5.0g NaCl, 20g agar, pH 7.2.
(3) Culture medium of autotoxic substance degrading bacteria: NH per liter4NO3 1.0g，NaCl 0.5g，K2HPO4·3H2O 1.5g，MgSO4·7H2O 0.5g，(NH4)2SO40.5g, pH 7.2. Weighing the components, dissolving the components in 1000mL of distilled water, sterilizing the solution for 15min at 121 ℃ under high pressure, cooling the solution to 45-50 ℃, and adding the solutionVarious autotoxic substances (gallic acid, coumaric acid, 3, 4-dihydroxy benzoic acid, p-hydroxybenzoic acid, vanillic acid, syringic acid, vanillin, ferulic acid and benzoic acid) identified in the continuous cropping soil of the heterophylly falsestarwort root are mixed with the total concentration of the phenolic acid of 1000mg/L, and after 10 percent of heterophylly falsestarwort root rhizosphere soil suspension is inoculated for enrichment culture for several generations, beneficial bacteria with autotoxic substance degradation capability and antagonistic action on pathogenic bacteria of the heterophylly falsestarwort root are screened.
Screening and identifying to obtain 12 strains of radix Pseudostellariae rhizosphere indigenous beneficial bacteria with strong antagonistic capability, including 2 strains of Burkholderia plantarfaria and Burkholderia cepacia; 4 strains of Pseudomonas (Pseudomonas fluorescens, Pseudomonas putida, Pseudomonas protegens, Pseudomonas korea); 6 strains of Bacillus (Bacillus amyloliquefaciens, Bacillus subtilis, Bacillus cereus, Bacillus pumilus, Bacillus megaterium, Bacillus mycoides). Wherein the strong antagonistic bacteria of Burkholderia cepacia, Burkholderia ambifaria, Pseudomonas protegens and Pseudomonas korea have been deposited in the general microbiological center of China Committee for culture Collection of microorganisms at 2016, 1 month and 28 days, and the deposit numbers are respectively: CGMCC NO.12119, CGMCC NO.12120, CGMCC NO.12121, CGMCC NO. 12122.
The indigenous antagonistic bacteria of the invention are obtained by a large amount of screening work, are obtained by screening 3000 soil bacteria, and have strong antagonistic effect on the special pathogenic bacteria of radix pseudostellariae (such as fusarium oxysporum, fusarium moniliforme, talaromyces tarda, etc.), as shown in fig. 2. The indigenous antagonistic bacteria disclosed by the invention not only can inhibit pathogenic bacteria, but also has the phosphate solubilizing capability, can effectively activate soil nutrients, and has important effects on improving the physicochemical properties of soil and promoting the absorption of plant nutrients as shown in figure 3.
Example 2 preparation of the Special Complex microbial Agents
The special compound microbial agent is prepared from general beneficial bacteria and indigenous beneficial bacteria in a volume ratio of 4:6, and the specific preparation method is as follows:
2.1 preparation of EM composite bacterial liquid
Adding 300g of brown sugar and 50mL of MS mother liquor into 18L of well water (tap water is used after being placed for at least 2 days), adding 1L of activated EM liquid strain, placing the well water in a room at 35-37 ℃ for closed fermentation for 2-3 weeks, opening a closed container opening every other day from the second day of fermentation to deflate and decompress once, and smelling sour and sweet taste when opening the container opening, wherein the normal phenomenon is that the fermentation expanding culture is successful and is reserved. The successful finished product fermentation liquor has acid and sweet taste, the pH value is 3.0-4.0, and the viable count is more than 100 hundred million/mL.
2.2 activation and expansion culture of indigenous beneficial bacteria
1) Strain activation
Respectively inoculating Pseudomonas fluorescens, Pseudomonas putida, Pseudomonas protegens and Pseudomonas korea in a Pseudomonas selective solid culture medium, and placing the culture medium in an incubator at the temperature of 28-32 ℃ for activation for 12-16 h.
Respectively inoculating other indigenous beneficial bacteria (including Burkholderia and bacillus) into LB solid culture, and activating in an incubator at 37 ℃ for 8-16 h.
2) Expanding culture
Inoculating activated various indigenous beneficial bacteria into liquid culture for fermentation culture, performing amplification culture on pseudomonas at 28-32 ℃, performing amplification culture on other indigenous beneficial bacteria at 37 ℃, and allowing the number of viable bacteria to be higher than 1.0 × 109Stopping the amplification culture at CFU/mL for later use.
2.3 preparation of Complex microbial Agents
Mixing the indigenous beneficial bacteria, namely Burkholderia, pseudomonas and bacillus according to the volume ratio of 4:1:1 to prepare the indigenous beneficial bacteria composite bacterial liquid (IM composite bacterial liquid).
Further, the EM composite bacterial liquid and the IM composite bacterial liquid are mixed according to the volume ratio of 4:6 to prepare the special composite microbial agent for the radix pseudostellariae.
Example 3 evaluation of field effect of special compound microbial inoculant for preventing and treating radix pseudostellariae continuous cropping obstacle
Firstly, fermenting the prepared special compound microbial inoculum and organic materials together to prepare a biological organic fertilizer, using the biological organic fertilizer as a base fertilizer to improve the continuous cropping soil of the radix pseudostellariae in advance, then preparing soil and ridging, planting the radix pseudostellariae, irrigating a proper amount of special compound microbial liquid inoculum from the seedling stage of the radix pseudostellariae in time, and performing yield difference comparison in the harvest stage. The concrete control measures are as follows:
3.1 preparation of microbial organic base fertilizer
Mixing and uniformly stirring organic materials of bean cypress, rice bran, dried pig manure, fish meal and brown sugar according to the weight ratio of 2:1:1:1:0.03, adding a proper amount of special composite microbial liquid fungicide, adjusting the amount of the added composite microbial liquid fungicide according to the dryness of the materials, controlling the water content to be 30% (preferably, the materials are easy to agglomerate but do not produce water when being kneaded by hands), fully stirring uniformly, bagging or barreling for closed anaerobic fermentation, carrying out fermentation at 37 ℃ for 48 hours, standing at room temperature for 2-4 weeks, and then ripening. And (3) inoculating a proper amount of special microbial liquid microbial inoculum into the anaerobic fermentation product, and fermenting for 48 hours at 37 ℃ in closed and ventilated equipment to obtain the special microbial organic fertilizer, wherein the total viable count is 1.0-5.0 hundred million/g.
3.2 soil improvement
Treatment group: after harvesting the previous crops, performing first soil preparation, applying 1200kg of microbial organic fertilizer which is fermented in advance (fermentation is started at least 1 month earlier) to each mu, deeply ploughing and turning into soil, uniformly raking up furrows, covering with black mulching films, irrigating furrows with feathered water until the surfaces of the furrows are slightly wet (but not submerged to the surfaces of the furrows), stopping water supply, and covering with films for fermentation for at least 1 month to improve and repair the deteriorated continuous cropping soil environment.
Control group: and (3) applying the same amount of base materials for fermenting the microbial organic fertilizer, not applying a special microbial liquid microbial inoculum in the later period, and keeping other field management measures consistent with the treatment group.
Blank group: the method is not applied to the bottom materials of the fermented microbial organic fertilizer, and special microbial liquid inoculants are not applied in the later period, so that other field management measures are consistent with those of a treatment group.
3.3 planting of Pseudostellaria heterophylla
And (3) performing soil preparation for the second time one week before planting, and opening straight-row grooves with the depth of 7-10 cm on the prepared furrow surfaces according to the row spacing of 15 cm. Before planting, 20kg of compound fertilizer, 25kg of phosphate fertilizer and 10kg of potash fertilizer are uniformly mixed per mu and then applied to planting furrows as seed fertilizers, a little amount of covering soil is added, then the seed seeds are obliquely arranged on the walls of the furrows according to the plant spacing of 5-7 cm, the bud heads are upward, the bud spacing is about 5cm from the horizon, and the soil is lightly covered. Planting 25-35 kg of seed seeds per mu from 11 late ten days to 12 months.
3.4 additional bacterium solution
In the middle and late ten days of 2 months, after the seedlings come out of the soil, 100 times of diluted special compound microbial inoculum is applied to the roots for 1 time, in the middle of 3 months and in the middle of 4 months, and 50 times of diluted special compound microbial inoculum is applied to the roots respectively for 1 time so as to resist the occurrence of soil-borne diseases. The root irrigation times are adjusted according to the growth vigor and the morbidity of the radix pseudostellariae, and can be properly increased if the morbidity symptom exists. Equal amount of water was applied to the control and blank groups.
3.5 Harvest and yield Difference comparison
When the plant of the radix pseudostellariae withers and falls into seedlings in the last ten days of 6 months, the radix pseudostellariae is harvested on sunny days. Digging more than 15 cm deeply with a small hoe, completely digging out radix Pseudostellariae root tuber to avoid damaging root tuber, picking out, cleaning, air drying, and weighing fresh weight.
Years of test results show that compared with the continuous cropping radix pseudostellariae (a control group and a blank group), the special microbial agent applied can obviously inhibit the morbidity, improve the acre yield of the radix pseudostellariae and increase the yield by about 30-65%. Therefore, the special microbial agent disclosed by the invention has an obvious field effect on prevention and treatment of the continuous cropping obstacles of the radix pseudostellariae, is stable in effect and has a huge market application prospect.
All modifications which can be derived or suggested by a person skilled in the art from the present disclosure are to be considered within the scope of the invention.
Claims (5)
1. A special compound microbial agent for eliminating successive cropping obstacles of radix pseudostellariae is characterized in that: the microbial inoculum consists of general beneficial bacteria and indigenous beneficial bacteria according to the volume ratio of 4: 6; the general beneficial bacteria are prepared by mixing lactic acid bacteria, photosynthetic bacteria, saccharomycetes and actinomycetes according to the volume ratio of 1:1:1: 1; the indigenous beneficial bacteria are formed by mixing Burkholderia, Pseudomonas and bacillus according to the volume ratio of 4:1:1, wherein the Burkholderia is one or two of Burkholderia ambifaria and Burkholderia cepacia, the Pseudomonas is one or more of Pseudomonas fluorescens, Pseudomonas putida, Pseudomonas protegens and Pseudomonas koreana, the bacillus is one or more of Bacillus amyloliquefaciens, Bacillus subtilis, Bacillus cereus, Bacillus pumilus, Bacillus megaterium and Bacillus mycoides, the preservation numbers of Burkholderia ambifaria and Burkholderia cepacia are respectively as follows: CGMCC NO.12120, CGMCC NO.12119, Pseudomonas protegens and Pseudomonas korea Korea have the preservation numbers of: CGMCC NO.12121 and CGMCC NO. 12122.
2. The special compound microbial inoculant for eliminating successive cropping obstacles of radix pseudostellariae as claimed in claim 1, wherein: the lactobacillus is one or more of lactobacillus plantarum, streptococcus lactis and lactobacillus casei.
3. The special compound microbial inoculant for eliminating successive cropping obstacles of radix pseudostellariae as claimed in claim 1, wherein: the photosynthetic bacteria are one or two of rhodopseudomonas palustris and rhodobacter sphaeroides.
4. The special compound microbial inoculant for eliminating successive cropping obstacles of radix pseudostellariae as claimed in claim 1, wherein: the yeast is one or two of saccharomyces cerevisiae and candida utilis.
5. The special compound microbial inoculant for eliminating successive cropping obstacles of radix pseudostellariae as claimed in claim 1, wherein: the actinomycetes are one or two of streptomyces griseus and streptomyces albus.
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| CN106433676A (en) * | 2016-09-23 | 2017-02-22 | 河海大学 | Soil conditioner and application thereof in organic agriculture fertilizing for newly cultivated coastal beach saline-alkali soil |
| CN107118997B (en) * | 2017-07-11 | 2019-11-15 | 福建贝迪药业有限公司 | A kind of feeding clostridium butyricum, the probiotics for promoting growth of animals or poultry and preparation method thereof |
| CN109497089A (en) * | 2019-01-22 | 2019-03-22 | 山东土木启生物科技有限公司 | With the complex micro organism fungicide and preparation method for reducing continuous cropping, wilt disease |
| CN111096336A (en) * | 2019-12-13 | 2020-05-05 | 广东新景象生物工程有限公司 | Preparation for preventing and treating crop native disease and preparation method thereof |
| CN111286480B (en) * | 2020-03-31 | 2022-04-12 | 中国科学院微生物研究所 | Compound microbial agent and application thereof in prevention and treatment of various plant diseases |
| CN112708572B (en) * | 2020-07-17 | 2023-10-24 | 兰州理工大学 | Microbial compound bacterial agent for improving yield and quality of dangshen, preparation method and application |
| CN112266881B (en) * | 2020-10-20 | 2021-11-19 | 山东农业大学 | Bacillus amyloliquefaciens and application thereof in preventing and treating apple continuous cropping obstacle |
| CN112314626B (en) * | 2020-11-09 | 2022-04-01 | 青岛农业大学 | Application of biological stimulin for improving activity of continuous cropping salvia miltiorrhiza seedlings |
| CN113099996A (en) * | 2021-04-21 | 2021-07-13 | 河南农业大学 | Method for eliminating continuous cropping obstacles of rehmannia and application |
| CN113684148B (en) * | 2021-08-19 | 2023-04-14 | 云南中烟工业有限责任公司 | Burkholderia cepacia, and isolated culture method and application thereof |
| CN114480203A (en) * | 2022-02-18 | 2022-05-13 | 北京意科乐生态科技有限公司 | Composite microbial preparation for resisting continuous cropping obstacles of Chinese herbal medicines and application method |
| CN114868773B (en) * | 2022-03-24 | 2024-03-29 | 中诚国联（河南）生物科技有限公司 | Compound microbial agent for preventing and treating anthracnose of fruit trees as well as preparation method and application thereof |
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