CN117187077A - Composite flora and application thereof, and control method of bacterial wilt of plants - Google Patents
Composite flora and application thereof, and control method of bacterial wilt of plants Download PDFInfo
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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
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- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
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Abstract
The invention discloses a compound flora and application thereof, and a method for preventing and treating plant bacterial wilt, wherein the compound flora comprises basket bacteria F34, aspergillus awamori F001 and methylobacterium YT2019B002. In the invention, the basket bacteria with the antagonism of the bacterial wilt, the aspergillus awamori with the phosphorus dissolving effect and the methylobacterium with the growth promoting effect are constructed as the compound bacterial group, and the bacterial group has a stable control effect on the bacterial wilt of tomatoes, can effectively delay the occurrence of the bacterial wilt, can effectively promote the improvement of the biomass of tomatoes, and has important significance in the aspect of controlling agricultural biological diseases.
Description
Technical Field
The invention belongs to the technical field of biological control of agricultural diseases, and in particular relates to a composite flora and application thereof in controlling or improving plant biomass, a control method of the plant bacterial wilt and a method for improving the plant biomass.
Background
Tomato is one of important crops in China, and has higher edible value and good cultivation and planting prospect as a fruit and vegetable co-feeding crop. Bacterial wilt of tomatoes is an important soil-borne disease in a tomato planting process, is caused by infection of Lawsonia of the Solanaceae (Ralstonia solanacearum) through wounds or natural orifices of tomatoes, has wide occurrence area and serious harm, and causes huge economic loss to tomato planting. Therefore, the prevention and treatment work of the bacterial wilt is particularly important for tomato planting production. Chemical control is still the most main control means of tomato bacterial wilt at present, but a large amount of pesticide and fertilizer inputs cause unbalance of soil beneficial microorganism communities and increasing rampant of harmful pathogenic bacteria, and simultaneously bring about a plurality of problems of environmental pollution, pesticide residues and the like.
Biological control by utilizing beneficial microorganisms is a novel way of controlling plant diseases which is safe, environment-friendly and has potential. Currently, biological control studies on the strain of the genus Laurella (R.solanacearum) are continuously carried out worldwide, and studies found from literature data show that a large number of antagonistic strains including fungi, bacteria, actinomycetes and the like are separated and screened from different materials, and the strains have good capability of inhibiting the growth of the bacterial wilt on a flat plate, and even though strains of the same species are screened, the biological control efficiency on the bacterial wilt of tomatoes is greatly different in potting experiments due to different material sources or habitats of the strains.
A plurality of researches show that the single antagonistic bacteria has good capability of inhibiting the growth of the bacterial wilt under the in-vitro condition, but after the single antagonistic bacteria are applied to the rhizosphere soil environment of tomatoes, the biological activity of the single antagonistic bacteria can be inhibited, so that the antagonistic capability is unstable and the like, and further the control effect on the bacterial wilt is poor. Therefore, the single antagonistic bacteria and other functional strains are compounded to construct a compound bacterial group with more stable functionality, which is more beneficial to healthy growth of plants and balance of a soil micro-ecological system.
Disclosure of Invention
Based on the above, the present invention aims to provide a complex bacterial flora and application thereof, and a method for preventing and treating plant bacterial wilt and a method for improving plant biomass.
The technical scheme for realizing the aim of the invention comprises the following steps.
In a first aspect of the invention, there is provided a complex flora comprising a basket F34, an Aspergillus awamori F001 and a Methylobacillus YT2019B002; the preservation number of the basket bacteria F34 is GDMCCNo.63413; the preservation number of the aspergillus awamori F001 is GDMCC No.63412; the deposit number of the methyl bacillus YT2019B002 is GDMCC No.61052.
In a second aspect, the invention provides the use of the complex bacterial flora in the control of bacterial wilt in plants or in the improvement of plant biomass.
In a third aspect of the present invention, there is provided a method for controlling bacterial wilt of plants, comprising the steps of: the complex flora is irrigated in the culture medium of tomatoes.
In a fourth aspect of the invention, there is provided a method of increasing plant biomass comprising the steps of: the complex flora is irrigated in the culture medium of tomatoes.
The antagonistic strain of the invention, basket (Talaromyces sp.) F34, is classified and named as Talaromyces sp, and is deposited in the microorganism strain collection of Guangdong province at the month 27 of 2023, with the deposit number of GDMCC No.63413 and the deposit unit address: guangzhou City first middle road No. 100 college No. 59 building No. 5 Guangdong province microbiological institute.
The phosphorus-dissolving strain Aspergillus awamori (Aspergillus awamori) F001 provided by the invention is classified and named Aspergillus awamori, and is preserved in the microorganism strain preservation center of Guangdong province at the year 2023, month 4 and 27, and the preservation number is GDMCC No.63412, and the preservation unit address is: guangzhou City first middle road No. 100 college No. 59 building No. 5 Guangdong province microbiological institute.
The growth-promoting strain methyl bacillus (methyl bacteria populi) YT2019B002 is classified and named as methyl bacteria populi, and is preserved in the Guangdong province microorganism strain preservation center on 6 th and 9 th of 2020, with the preservation number of GDMCC No.61052 and the preservation unit address: guangzhou City first middle road No. 100 college No. 59 building No. 5 Guangdong province microbiological institute.
In the invention, the basket bacteria with the antagonism of the bacterial wilt, the aspergillus awamori with the phosphorus dissolving effect and the methylobacterium with the growth promoting effect are constructed as the compound bacterial group, and the bacterial group has a stable control effect on the bacterial wilt of tomatoes, can effectively delay the occurrence of the bacterial wilt, can effectively promote the improvement of the biomass of tomatoes, and has important significance in the aspect of controlling agricultural biological diseases.
Drawings
FIG. 1 shows the results of the measurement of the affinity between strains in example 1 of the present invention.
FIG. 2 shows the results of a potted plant control experiment of tomato bacterial wilt by a single strain and a complex bacterial population in example 2 of the present invention.
FIG. 3 shows the effect of each treatment group on tomato growth without receiving bacterial wilt in example 3 of the present invention.
Detailed Description
The present invention will be described more fully hereinafter in order to facilitate an understanding of the present invention. This invention may be embodied in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The term "and/or" as used herein includes any and all combinations of one or more of the associated listed items.
The experimental procedures, which do not address the specific conditions in the examples below, are generally followed by conventional conditions, such as those described in Green and Sambrook et al, molecular cloning, an experimental guideline (Molecular Cloning: A Laboratory Manual, 2013), or by the manufacturer's recommendations. The various chemicals commonly used in the examples are commercially available.
In the invention, firstly, a composite bacterial colony with functions of resisting bacterial wilt, dissolving phosphorus, promoting growth and the like is constructed through an affinity test on a basket bacterial F34 with the antagonism of the bacterial wilt, an aspergillus awamori F001 with the action of dissolving phosphorus and a methyl bacillus YT2019B002 bacterial strain with the action of promoting growth, and then the composite bacterial colony is found to have stable prevention and treatment effects on tomato bacterial wilt through a pot experiment, so that the bacterial wilt can be effectively delayed (the phenomenon that the tomato is infected by the bacterial wilt and the bacterial wilt is developed after the bacterial wilt is inoculated for 14d, the disease index of 21d after inoculation is only 25.0 percent, and the prevention and treatment effects reach 70.71 percent). And can effectively promote the improvement of tomato biomass (compared with a control, the compound flora can effectively promote the growth of tomatoes, and obviously improve the growth indexes such as fresh weight, dry weight, stem length, stem diameter, chlorophyll content and the like), and has important significance in the aspect of preventing and controlling agricultural biological diseases.
In some embodiments of the invention, a complex flora is disclosed, comprising a basket F34, a. Awamori F001, and a bacillus methylobacterium YT2019B002; the preservation number of the basket bacteria F34 is GDMCC No.63413; the preservation number of the aspergillus awamori F001 is GDMCC No.63412; the deposit number of the methyl bacillus YT2019B002 is GDMCC No.61052.
The nucleotide sequence of basket F34 is as follows (SEQ ID NO. 1): TGGAAGTAAAAGTCGTAACAAGGTTTCCGTAGGTGAACCTGCGGAAGGATCATTACTGAGTGAGGGCCCCTCGGGGTCCAACCTCCCACCCGTGTTTAACGAACCTTTGTTGCTTCGGCGGGCCCGCCTCACGGCCGCCGGGGGGCTCCTGCCCCCGGGCCCGCGCCCGCCGAAGCCCCCCCTTGAACGCTGTCTGAAGTTTGCAGTCTGAGAAACTAGCTAAATTAGTTAAAACTTTCAACAACGGATCTCTTGGTTCCGGCATCGATGAAGAACGCAGCGAAATGCGATAACTAATGTGAATTGCAGAATTCAGTGAATCATCGAGTCTTTGAACGCACATTGCGCCCTCTGGTATTCCGGAGGGCATGCCTGTCCGAGCGTCATTGCTGCCCTCAAGCACGGCTTGTGTGTTGGGCCCCCGTCCCCCCCTCTGCCGGGGGGACGGGCCCGAAAGGCAGCGGCGGCACCGCGTCCGGTCCTCGAGCGTATGGGGCTTCGTCACCCGCTCTTGTAGGCCCGGCCGGCGCCAGCCGACCCCAACCCTAAATTTTTATCAGGTTGACCTCGGATCAGGTACGGA
The nucleotide sequence of aspergillus awamori F001 has been submitted to GenBank under the number MT953924.
The nucleotide sequence of methylobacterium YT2019B002 has been submitted to GenBank under the number MT568576.
In some of these embodiments, the complex flora consists of basket F34, aspergillus awamori F001, and methylobacterium YT2019B002.
In some embodiments, the concentration of the basket F34 in the complex population is 0.8X10 8 ～1.2×10 8 spores/g; the concentration of Aspergillus awamori F001 is 0.8X10 5 ～1.2×10 5 spores/g; the concentration of the methyl bacillus YT2019B002 is 0.8X10 8 ～1.2×10 8 cfu/g。
In some embodiments, the concentration of the basket F34 in the complex population is 0.9X10 8 ～1.1×10 8 spores/g; the concentration of Aspergillus awamori F001 is 0.9X10 5 ～1.1×10 5 spores/g; the concentration of the methyl bacillus YT2019B002 is 0.9X10 8 ～1.1×10 8 cfu/g。
In some embodiments, the concentration of the basket F34 in the complex population is 0.95X10 8 ～1.05×10 8 spores/g; the concentration of Aspergillus awamori F001 is 0.95X10 5 ～1.05×10 5 spores/g; the concentration of the methyl bacillus YT2019B002 is 0.95 multiplied by 10 8 ～1.05×10 8 cfu/g。
In other embodiments of the present invention, the use of the complex bacterial flora described above for controlling bacterial wilt in plants is disclosed.
In some embodiments, the plant is tomato.
In other embodiments of the present invention, a method for controlling tomato bacterial wilt is disclosed, comprising the steps of: the complex flora is irrigated in the culture medium of tomatoes.
In some embodiments, the concentration of the basket F34 in the culture medium is 0.8X10 8 ～1.2×10 8 spores/g; aspergillus awamori F001 at a concentration of 0.8X10 5 ～1.2×10 5 spores/g; the concentration of Methylobacillus YT2019B002 was 0.8X10 8 ～1.2×10 8 cfu/g。
In some embodiments, the concentration of the basket F34 in the culture medium is 0.9X10 8 ～1.1×10 8 spores/g; the concentration of Aspergillus awamori F001 was 0.9X10 5 ～1.1×10 5 spores/g; the concentration of Methylobacillus YT2019B002 was 0.9X10 8 ～1.1×10 8 cfu/g。
In some embodiments, the concentration of the basket F34 in the culture medium is 0.95X10 8 ～1.05×10 8 spores/g; the concentration of Aspergillus awamori F001 was 0.95X10 5 ～1.05×10 5 spores/g; the concentration of Methylobacillus YT2019B002 was 0.95X10 8 ～1.05×10 8 cfu/g。
In other embodiments of the present invention, the use of the complex bacterial population described above for increasing plant biomass is disclosed.
In some embodiments, the plant is tomato.
In other embodiments of the present invention, a method of increasing tomato biomass is disclosed comprising the steps of: the complex flora is irrigated in the culture medium of tomatoes.
In some embodiments, the concentration of the basket F34 in the culture medium is 0.8X10 8 ～1.2×10 8 spores/g; aspergillus awamori F001 at a concentration of 0.8X10 5 ～1.2×10 5 spores/g; the concentration of Methylobacillus YT2019B002 was 0.8X10 8 ～1.2×10 8 cfu/g。
In some embodiments, the concentration of the basket F34 in the culture medium is 0.9X10 8 ～1.1×10 8 spores/g; the concentration of Aspergillus awamori F001 was 0.9X10 5 ～1.1×10 5 spores/g; the concentration of Methylobacillus YT2019B002 was 0.9X10 8 ～1.1×10 8 cfu/g。
In some embodiments, the concentration of the basket F34 in the culture medium is 0.95X10 8 ～1.05×10 8 spores/g; the concentration of Aspergillus awamori F001 was 0.95X10 5 ～1.05×10 5 spores/g; the concentration of Methylobacillus YT2019B002 was 0.95X10 8 ～1.05×10 8 cfu/g。
In the following examples, PDA medium (product No. 021050) and NA (product No. 022020) were purchased from Guangdong Cryptographic microorganism Co.
The invention is described in detail below with reference to the drawings and the specific embodiments.
Example 1 measurement of the affinity between strains
The functional strain to be tested, namely antagonistic strain basket bacteria (Talaromyces sp.) F34, phosphorus-dissolving strain Aspergillus awamori (Aspergillus awamori) F001 and growth-promoting strain methylobacterium (methylobacterium sporuli) YT2019B002 are respectively subjected to purification culture.
Beating a culture medium with hyphae into a bacterial cake with the diameter of 5mm by using a puncher, uniformly mixing a PDA culture medium and an NA culture medium according to equal proportion, pouring the culture medium, inoculating the bacterial cake into a culture dish after the culture medium is solidified, and carrying out pairwise confrontation with a distance of 2cm; after streaking the bacteria on the medium, the fungi were inoculated in the medium with a spacing of 2cm between the fungi and the bacterial lines, and the affinity between the bacteria and the fungi was observed. Each treatment is repeated for 3 times, the culture is inverted for 3 to 5 days at the constant temperature of 28 ℃, the mutual inhibition growth phenomenon is observed between each colony, and the generation of a bacteria inhibition zone indicates that the bacteria inhibition zone does not have affinity between the bacteria inhibition zone and the bacteria inhibition zone, and the complex flora cannot be built by compounding.
The results are shown in FIG. 1, and it can be seen from FIG. 1 that antagonistic bacteria are basket (Talaromyces sp.) F34, phosphorus-dissolving strain Aspergillus awamori (Aspergillus awamori) F001 and growth-promoting strain methyl bacillus YT2019B002,3 strains can normally grow on the same plate without mutual interference and mutual inhibition phenomenon, and have affinity between every two, which means that the three have a mixed condition, and a composite flora can be constructed together.
Example 2 control Effect of Compound flora on tomato bacterial wilt
1. Preparation of inoculant suspension
1. Bacterial wilt-inducing tomato plant stem isolated from solanacearum, genBank No. OR 342535) and methylobacterium prokinensis YT2019B002 are activated and cultured in NA culture medium in advance, single colony is selected and inoculated in LB liquid culture medium, shake-cultured at 30 deg.C and 180r/min to logarithmic growth phase, bacterial liquid is discarded and bacterial body is collected, bacterial wilt-inducing tomato plant stem and functional bacterial YT2019B002 suspension are prepared by using sterile bacteria respectively for standby.
2. Respectively inoculating phosphorus-dissolving bacteria F001 and antagonistic bacteria F34 into PDB liquid for culturing, oscillating at 30deg.C and 180r/min to logarithmic phase, centrifuging 10000g to remove bacterial liquid, collecting bacterial cells, re-suspending with sterile water containing 0.1% Tween-80, and preparing spore suspension for use.
2. Potted plant treatment
Transplanting tomato seedlings growing to 4-leaf stage into flowerpots with diameter and height of 10cm, uniformly mixing 100g of culture medium (comprising turfy soil and vermiculite in equal proportion before use, sterilizing for later use, and purchasing from Guangzhou large-scale agriculture science and technology Co., ltd., and measuringThe fixed component content is 0.3% of total nitrogen, 0.1% of total phosphorus and 93.2% of organic matter, wherein the content of effective phosphorus is 2.991 g/kg), and the bacterial wilt and the functional bacterial suspension are irrigated once at the 1 st d after the plant is fixed root, so that the bacterial wilt concentration in the culture medium is 1.0x10 9 cfu/g, methylobacillus YT2019B002 final concentration of 1.0X10 8 cfu/g, final concentration of lysophosphate F001 of 1.0X10 5 Spores/g final concentration of antagonistic F34 of 1.0X10 8 spores/g。
Before treatment, each tomato is subjected to root injury treatment by a sterile needle head, and the later-stage water and fertilizer management is consistent.
The test was designed as 10 treatments (as in Table 1), each treatment was repeated 3 times for 5 plants, and incubated in a climatic chamber (16 h light at 30 ℃ C.; 8h dark at 28 ℃ C.).
TABLE 1
3. Disease prevention index determination
The tomato bacterial wilt morbidity, disease index and prevention and treatment effect are statistically analyzed by continuously observing the functional bacteria and pathogenic bacteria inoculated to the tomato for 21d after treatment.
The bacterial wilt disease grade classification criteria are shown in table 2.
TABLE 2
| Disease grade | Degree of occurrence of disease |
| Level 0 | Healthy plants, without wilting symptoms |
| Level 1 | 1/4 leaf wilting |
| Level 2 | 1/2 leaf wilting |
| 3 grade | 2/3 leaf wilting |
| Grade 4 | Withering and death of whole plant |
4. Experimental results
The results of the control effect of single strain and compound flora treatment on tomato bacterial wilt are shown in table 3 and figure 2.
TABLE 3 Table 3
| Treatment of | Incidence (%) | Index of disease (%) | Preventing and curing effectFruit (%) |
| CK+RS | 100.00±0a | 80.56±9.62a | |
| T2 | 91.67±14.43a | 56.94±6.37a | 29.12±6.03b |
| T4 | 83.33±14.43a | 54.25±9.41ab | 31.22±18.47b |
| T6 | 75.00±25.00a | 54.33±20.20ab | 33.44±21.92ab |
| T8 | 33.33±25.00b | 25.00±25.00b | 70.71±27.50a |
Note that: the data in the table are mean ± standard deviation, the different lowercase letters after the same column of data represent the significance of the differences (P <0.05, lsd)
The results in Table 3 and FIG. 2 show that the control group CK+RS and the single strain treatment groups T2, T4 and T6 begin to develop after the 4 th d after inoculation of pathogenic bacteria, the disease index gradually rises, the incidence of the control group CK+RS reaches the highest value of 100% at 14d, and the developed plants appear to gradually wilt and die finally.
Compared with CK+RS, the incidence of tomato bacterial wilt is reduced to different degrees after the tomato is treated by functional bacteria, the disease development of the tomato bacterial wilt in a single strain treatment group or a compound bacterial group treatment group is delayed to a certain extent, the control effect is in the range of 29% -71%, compared with the single strain treatment, the disease time of the bacterial wilt is obviously delayed after the compound bacterial group treatment, the phenomenon that the tomato is infected with the bacterial wilt and is diseased after 14d of inoculation pathogenic bacteria is gradually discovered, the disease index is only 25.00% after 21d of inoculation, and the control effect reaches 70.71%.
Example 3 Effect of Complex flora on tomato growth
1. The preparation method of the bacterial suspension and the potting treatment method are the same as in test example 2.
2. And (3) continuously observing the tomato inoculated functional bacteria and pathogenic bacteria for 21d after treatment, and statistically analyzing indexes such as total fresh weight, total dry weight, stem length, stem diameter, chlorophyll content and the like related to plant growth.
3. The effect of single strain and complex flora on tomato growth without bacterial wilt is shown in fig. 3 and table 4, and the effect of single strain and complex flora on tomato growth without bacterial wilt is shown in table 4.
TABLE 4 Effect of single strains and Complex populations on tomato growth
Note that: the data in the table are mean ± standard deviation, the different lowercase letters after the same column of data represent the significance of the differences (P <0.05, lsd)
In the treatment group without bacterial wilt, the single strain treatment groups T1, T3 and T5 and the complex bacterial group treatment group T7 each significantly increased tomato biomass (fig. 3, including total fresh weight, total dry weight, stem length, stem diameter and chlorophyll content) compared to the blank group (CK). The total fresh weight was increased by 103.32%, 214.52%, 121.58% and 304.15%, the total dry weight was increased by 21.05%, 97.37%, 50.00% and 263.16%, the stem length was increased by 84.91%, 127.43%, 58.29% and 177.71%, the stem diameter was increased by 21.31%, 34.02%, 27.15% and 32.99%, and the chlorophyll content was also increased by 27.46%, 5.18%, 51.81% and 85.49%, respectively (table 4); comparison between single strain treatment groups, overall data were observed and comprehensive analysis showed that treatment group T3 had better effect on increasing tomato biomass than the other two single strain treatment groups T1 and T5, but overall complex flora treatment group T7 was more able to significantly increase accumulation of tomato biomass (fig. 3 and table 4).
As can be seen from the results in Table 4, in the treatment group inoculated with both functional bacteria and bacterial wilt, tomato plants were weakened and could not normally photosynthesis and biomass accumulated due to the invasion of bacterial wilt, but tomato growth conditions of the composite microbial inoculum treatment group T8 were superior to those of the control group CK+RS and the single strain treatment groups T2, T4 and T6, compared with (CK+RS), total fresh weights of the composite microbial inoculum treatment groups were increased by 271.68% (T2), 140.00% (T4), 281.82% (T6) and 311.76% (T8), total dry weights were increased by 655.56% (T2), 134.48% (T4), 183.33% (T6) and 223.81% (T8), stem lengths were increased by 45.86% (T2), 26.19% (T4), 41.45% (T6) and 52.37% (T8), respectively, and chlorophyll contents were increased by 13.89% (T2), 2.04% (T4), 3.07% (T6) and 48.01% (T8), respectively; compared with the single strain treatment groups (T2, T4 and T6), the growth indexes have no obvious difference, and the improvement of tomato biomass infected with bacterial wilt is not ideal. Under the infection of the bacterial wilt, compared with a single bacterial strain, the composite bacterial colony can also obviously improve the biomass of tomatoes and better exert the growth promoting effect.
The technical features of the above-described embodiments may be arbitrarily combined, and all possible combinations of the technical features in the above-described embodiments are not described for brevity of description, however, as long as there is no contradiction between the combinations of the technical features, they should be considered as the scope of the description.
The above examples illustrate only a few embodiments of the invention, which are described in detail and are not to be construed as limiting the scope of the invention. It should be noted that it will be apparent to those skilled in the art that several variations and modifications can be made without departing from the spirit of the invention, which are all within the scope of the invention. Accordingly, the scope of protection of the present invention is to be determined by the appended claims.
Claims (10)
1. A complex flora, wherein the complex flora comprises a basket fungus F34, an aspergillus awamori F001 and a methylobacterium YT2019B002; the preservation number of the basket bacteria F34 is GDMCCNo.63413; the collection number of the aspergillus awamori F001 is GDMCCNo.63412; the preservation number of the methylobacterium YT2019B002 is GDMCCNo.61052.
2. The complex of claim 1, wherein the complex is composed of a basket F34, a. Awamori F001, and a methylobacterium YT2019B002.
3. The complex of claim 1 or 2, wherein the concentration of the basket F34 in the complex is 0.8x10 8 ～1.2×10 8 spores/g; the concentration of Aspergillus awamori F001 is 0.8X10 5 ～1.2×10 5 spores/g; the concentration of the methyl bacillus YT2019B002 is 0.8X10 8 ～1.2×10 8 cfu/g；
Preferably, the concentration of the basket F34 in the complex bacterial group is 0.9X10 8 ～1.1×10 8 spores/g; the concentration of Aspergillus awamori F001 is 0.9X10 5 ～1.1×10 5 spores/g; the concentration of the methyl bacillus YT2019B002 is 0.9X10 8 ～1.1×10 8 cfu/g；
More preferably, the concentration of the basket F34 in the complex is 0.95X10 8 ～1.05×10 8 spores/g; the concentration of Aspergillus awamori F001 is 0.95X10 5 ～1.05×10 5 spores/g; the concentration of the methyl bacillus YT2019B002 is 0.95 multiplied by 10 8 ～1.05×10 8 cfu/g。
4. Use of a complex bacterial population according to any one of claims 1 to 3 for controlling bacterial wilt in plants.
5. The use according to claim 4, wherein the plant is tomato.
6. The control method of tomato bacterial wilt is characterized by comprising the following steps: the complex flora is irrigated in the culture medium of tomatoes.
7. The method for controlling bacterial wilt of tomato according to claim 6, wherein the concentration of the basket bacteria F34 in the culture medium is 0.8 x 10 8 ～1.2×10 8 spores/g; aspergillus awamori F001 at a concentration of 0.8X10 5 ～1.2×10 5 spores/g; the concentration of Methylobacillus YT2019B002 was 0.8X10 8 ～1.2×10 8 cfu/g；
Preferably, the concentration of the basket F34 in the culture medium is 0.9X10 8 ～1.1×10 8 spores/g; the concentration of Aspergillus awamori F001 was 0.9X10 5 ～1.1×10 5 spores/g; the concentration of Methylobacillus YT2019B002 was 0.9X10 8 ～1.1×10 8 cfu/g；
More preferably, the concentration of the basket F34 in the culture medium is 0.95X10 8 ～1.05×10 8 spores/g; the concentration of Aspergillus awamori F001 was 0.95X10 5 ～1.05×10 5 spores/g; the concentration of Methylobacillus YT2019B002 was 0.95X10 8 ～1.05×10 8 cfu/g。
8. Use of a complex bacterial population according to any one of claims 1 to 3 for increasing biomass in tomatoes.
9. A method for increasing biomass in tomatoes, comprising the steps of: the complex flora is irrigated in the culture medium of tomatoes.
10. A method for increasing tomato biomass according to claim 9, wherein the concentration of basket F34 in the culture medium is 0.8 x 10 8 ～1.2×10 8 spores/g; aspergillus awamori F001 at a concentration of 0.8X10 5 ～1.2×10 5 spores/g; the concentration of Methylobacillus YT2019B002 was 0.8X10 8 ～1.2×10 8 cfu/g；
Preferably, the concentration of the basket F34 in the culture medium is 0.9X10 8 ～1.1×10 8 spores/g; the concentration of Aspergillus awamori F001 was 0.9X10 5 ～1.1×10 5 spores/g; the concentration of Methylobacillus YT2019B002 was 0.9X10 8 ～1.1×10 8 cfu/g；
More preferably, the concentration of the basket F34 in the culture medium is 0.95X10 8 ～1.05×10 8 spores/g; the concentration of Aspergillus awamori F001 was 0.95X10 5 ～1.05×10 5 spores/g; the concentration of Methylobacillus YT2019B002 was 0.95X10 8 ～1.05×10 8 cfu/g。
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Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2006065985A2 (en) * | 2004-12-16 | 2006-06-22 | University Of Florida Research Foundation, Inc. | Materials and methods for the control of bacterial wilt in plants |
| CN106591157A (en) * | 2017-02-23 | 2017-04-26 | 北京市农林科学院 | Aspergillus tubingensis with disease prevention and growth promoting functions as well as preparation and application of aspergillus tubingensis metabolites |
| CN106635922A (en) * | 2017-01-22 | 2017-05-10 | 浙江农林大学 | Salt-tolerant biocontrol bacterium B268 for bacterial wilt of horsetail beefwood and application of salt-tolerant biocontrol bacterium B268 |
| CN108384734A (en) * | 2018-02-22 | 2018-08-10 | 广西壮族自治区农业科学院微生物研究所 | A kind of biocontrol agent and preparation method thereof of prevention tobacco rhizome class disease |
| CN108676741A (en) * | 2018-04-26 | 2018-10-19 | 南京农业大学 | A kind of functionality complex microorganism seedling medium and the preparation method and application thereof |
| CN109504622A (en) * | 2018-11-16 | 2019-03-22 | 广东植物龙生物技术股份有限公司 | A kind of lysine bacillus and the composite bacteria agent of bacillus subtilis and preparation method thereof |
| CN110300514A (en) * | 2017-01-03 | 2019-10-01 | 孟山都技术公司 | Microbial composite and method |
| CN115029276A (en) * | 2022-06-24 | 2022-09-09 | 中国烟草总公司四川省公司 | Acid-resistant antagonistic bacterium of ralstonia solanacearum and application thereof |
| CN115369062A (en) * | 2022-09-02 | 2022-11-22 | 上海市农业科学院 | Tomato bacterial wilt antagonistic bacterium WJB0802 and application thereof |
| CN115369064A (en) * | 2022-09-30 | 2022-11-22 | 海南师范大学 | Development and application of multifunctional composite microbial inoculum for enhancing rhizosphere colonization and control effect of biocontrol bacillus belgii |
-
2023
- 2023-07-24 CN CN202310911084.3A patent/CN117187077B/en active Active
Patent Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2006065985A2 (en) * | 2004-12-16 | 2006-06-22 | University Of Florida Research Foundation, Inc. | Materials and methods for the control of bacterial wilt in plants |
| CN110300514A (en) * | 2017-01-03 | 2019-10-01 | 孟山都技术公司 | Microbial composite and method |
| CN106635922A (en) * | 2017-01-22 | 2017-05-10 | 浙江农林大学 | Salt-tolerant biocontrol bacterium B268 for bacterial wilt of horsetail beefwood and application of salt-tolerant biocontrol bacterium B268 |
| CN106591157A (en) * | 2017-02-23 | 2017-04-26 | 北京市农林科学院 | Aspergillus tubingensis with disease prevention and growth promoting functions as well as preparation and application of aspergillus tubingensis metabolites |
| CN108384734A (en) * | 2018-02-22 | 2018-08-10 | 广西壮族自治区农业科学院微生物研究所 | A kind of biocontrol agent and preparation method thereof of prevention tobacco rhizome class disease |
| CN108676741A (en) * | 2018-04-26 | 2018-10-19 | 南京农业大学 | A kind of functionality complex microorganism seedling medium and the preparation method and application thereof |
| CN109504622A (en) * | 2018-11-16 | 2019-03-22 | 广东植物龙生物技术股份有限公司 | A kind of lysine bacillus and the composite bacteria agent of bacillus subtilis and preparation method thereof |
| CN115029276A (en) * | 2022-06-24 | 2022-09-09 | 中国烟草总公司四川省公司 | Acid-resistant antagonistic bacterium of ralstonia solanacearum and application thereof |
| CN115369062A (en) * | 2022-09-02 | 2022-11-22 | 上海市农业科学院 | Tomato bacterial wilt antagonistic bacterium WJB0802 and application thereof |
| CN115369064A (en) * | 2022-09-30 | 2022-11-22 | 海南师范大学 | Development and application of multifunctional composite microbial inoculum for enhancing rhizosphere colonization and control effect of biocontrol bacillus belgii |
Non-Patent Citations (4)
| Title |
|---|
| LOURENA ARONE MAXWELL等: "Evaluation of Biocontrol Agents Against Bacterial Wilt in Tomato Using Seedling Screening", PROCEEDINGS OF THE INTERNATIONAL SYMPOSIUM SOUTHEAST ASIA VEGETABLE 2021 (SEAVEG 2021) * |
| NEELU SINGH等: "Effects of Bacillus subtilis, Pseudomonas fluorescens and Aspergillus awamori on the wilt-leaf spot disease complex of tomato", PHYTOPARASITICA, vol. 43, pages 61 - 75 * |
| 李勤奋等: "复合菌剂对番茄抗病促生的效果研究", 生态环境学报, vol. 21, no. 11, pages 1836 - 1840 * |
| 谢华蓉等: "广藿香内生真菌多样性及其对青枯菌的拮抗活性", 微生物学通报, vol. 44, no. 05, pages 1171 - 1181 * |
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