CN112522146A - Growth-promoting disease-resistant bacillus amyloliquefaciens and application thereof - Google Patents
Growth-promoting disease-resistant bacillus amyloliquefaciens and application thereof Download PDFInfo
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- 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
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- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
- C05G3/60—Biocides or preservatives, e.g. disinfectants, pesticides or herbicides; Pest repellants or attractants
Abstract
The invention discloses a Bacillus amyloliquefaciens YQ1-2, which is Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) YQ1-2 with the preservation number as follows: CGMCC No. 20309. The invention also discloses the application of the bacillus amyloliquefaciens YQ 1-2: has growth promoting effect on plant, and can improve plant disease resistance.
Description
Technical Field
The invention belongs to the field of agricultural microorganisms, and particularly relates to bacillus amyloliquefaciens, a culture method thereof and application thereof in agricultural planting.
Background
The microbial diseases of crops are always the big problems troubling agricultural production, and are one of the main reasons causing the yield reduction of crops and influencing economic benefits. For a long time, the use of chemical pesticides is a main means for preventing and treating crop microbial diseases, but pesticide residues are caused, so that on one hand, the ecological environment is greatly influenced, and particularly, pathogenic microorganisms in a micro-ecological system generate drug resistance, so that the pesticide spraying concentration is higher and higher, the using amount is larger and larger, and the problem of pesticide residues is more and more serious. On the other hand, residual pesticides are transmitted hierarchically through a food chain, which brings potential risks to human health and has attracted high attention. In contrast, the biological control technology has the characteristics of safety, no toxicity, environmental protection and the like, and has remarkable advantages in the aspect of controlling vegetable diseases. Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) has been widely studied for its ability to inhibit plant diseases and promote plant growth. The compound has wide application prospect in the fields of biological control, environmental protection treatment, livestock product processing, biological fertilizer and the like.
Bacillus amyloliquefaciens can form spores resistant to high temperature, ultraviolet light, dryness, ionizing radiation and toxic chemicals under the stress of nutrient deficiency or other adverse conditions. The purpose of biological control is achieved by competing with pathogenic bacteria for nutrition and infection sites, secreting antibacterial substances and inducing plants to generate systemic disease resistance in the growth process. In addition, researches show that the bacillus amyloliquefaciens can also produce growth promotion effect on plants through different modes such as plant growth hormone secretion, phosphorus dissolving and nitrogen fixing effects, plant nutrition promotion functions and the like.
2016101744728 the invention, a peony endogenous Bacillus amyloliquefaciens, a separation method and an application thereof, tells that: GDMCC NO: 60008, the Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) Mdgb15 is an excellent plant endophytic disease-preventing growth-promoting bacterium, the fermentation liquid of the strain has good disease-preventing (peony gray mold) effect on potted peony, and simultaneously has good growth-promoting effect, the bio-control strain has the disease-preventing and growth-promoting effects, and through the application in peony greenhouse cultivation, the growth condition of the peony can be improved, and the disease incidence rate can be effectively controlled.
2017100361179 the invention of Bacillus amyloliquefaciens Y15 for preventing diseases, promoting growth and resisting stress of all plants and the application thereof reports that: the biological control effect of the bacillus amyloliquefaciens Y15 with the preservation number of CGMCC NO.13327 on various plant diseases (bacterial fruit blotch of watermelon, yellow leaf curl of tomato and peach leaf curl disease) after root irrigation treatment reaches more than 50 percent; the growth promoting effect on various crops reaches more than 18 percent; in the field, the control effect on various diseases is above 47.05%, and the growth promoting effect is very obvious; and can obviously improve the stress resistance of the plants.
2017103959466 the invention of Bacillus amyloliquefaciens 2YN11 aiming at the disease prevention, growth promotion, quality improvement and stress resistance of tomatoes and the application thereof reports that: the bio-control effect of the bacillus amyloliquefaciens 2YN11 with the preservation number of CGMCC NO.12606 on various tomato diseases (tomato bacterial wilt, tomato bacterial spot disease and tomato yellow leaf curl disease) after root irrigation treatment at least reaches more than 47.05 percent, and the effect is very obvious; the yield of the farmland crop treated by 2YN11 is increased by over 30.54 percent compared with a control group, the taste of the tomato is obviously improved, and various nutritional indexes are obviously improved.
Disclosure of Invention
The invention aims to solve the problem of providing a bacillus amyloliquefaciens and application thereof.
In order to solve the technical problems, the invention provides a Bacillus amyloliquefaciens YQ1-2, which is Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) YQ1-2, and the preservation number is as follows: CGMCC No. 20309.
The invention also provides the application of the bacillus amyloliquefaciens YQ 1-2: has growth promoting effect on plants.
As an improvement of the use of the Bacillus amyloliquefaciens YQ1-2 of the invention: improve the disease resistance of plants.
As a further improvement of the use of the Bacillus amyloliquefaciens YQ1-2 of the invention: the plant is a melon.
As a further improvement of the use of the Bacillus amyloliquefaciens YQ1-2 of the invention: has disease-resistant effect on the blight and root rot of sweet melons.
The root rot mainly infects roots and rhizomes, the infected parts are initially in a water stain shape and then in a tawny necrotic rot shape, white mold is generated on the surfaces of the rhizomes when the humidity is high, and vascular bundles at the rotten parts of the roots are browned but do not develop upwards, so that the root rot is obviously different from the blight.
The preservation information of the bacillus amyloliquefaciens YQ1-2 is as follows:
the preservation name is Bacillus amyloliquefaciens, and the preservation unit is as follows: china general microbiological culture Collection center, preservation Address: xilu No.1 Hospital No. 3, Beijing, Chaoyang, on Beijing, with a deposit number: CGMCC No.20309, preservation time 2020, 07/2020.
The bacillus amyloliquefaciens YQ1-2 is obtained by separating from dry kiln Zhejiang Jiaxing city dry-kiln Zhenzhu melon base; the colony morphology is characterized in that: the cells are rod-shaped, spore-forming, unsmooth in surface, white, irregular in edge and opaque.
The used fermentation medium is LB medium: 5g of yeast extract, 10g of tryptone, 10g of NaCl, and pH 7.0.
The Bacillus amyloliquefaciens YQ1-2 of the present invention also exerts an excellent effect in acidic (pH about 4.56) soil.
The bacillus amyloliquefaciens YQ1-2 has good safety, has the effects of promoting the growth of plants, improving the disease resistance and the like, and has wide application prospect in agricultural planting.
Drawings
The following describes embodiments of the present invention in further detail with reference to the accompanying drawings.
FIG. 1 is a photograph showing the form of cells;
FIG. 2 shows a micrograph of the strain (A) and spore morphology (B);
FIG. 3 shows the results of the plate confrontation test of the strain and Fusarium oxysporum;
FIG. 4 is a graph showing the effect of inoculum size on strain growth;
FIG. 5 is the effect of pH on strain growth;
FIG. 6 is the effect of NaCl concentration on strain growth;
FIG. 7 is the effect of temperature on strain growth;
FIG. 8 is the effect of temperature on sporulation of a strain;
figure 9 is the effect of different fertilization treatments on melon growth.
Detailed Description
The invention will be further described with reference to specific examples, but the scope of the invention is not limited thereto:
example 1:
(1) isolation of the Strain
The separation and screening method of the bacillus amyloliquefaciens at least comprises the following steps:
A. sampling:
collecting 5 bags of rhizosphere soil samples with good growth vigor from dry kiln Zhejiang Jiaxing city Jiashan county Zhenguang base, wherein the sampling time is 2018 years;
B. primary screening of strains:
the method for separating the bacillus amyloliquefaciens comprises the following steps: and (3) respectively treating each bag of soil sample as follows: weighing 5g of sample, placing in 45ml of sterile water, shaking for 30min on a shaking table, then sucking 0.5ml to 9.5ml of sterile water of the uniformly shaken soil solution, and diluting until 10-6The suction dilution factor is 10-4、10-5、10-60.1ml of each solution is coated on a NA solid medium plate; culturing the coated plate in an incubator at 30 ℃ for 24h, finally selecting 8 strains of different genera and fast growing strains according to the colony morphology difference, and further screening;
C. re-screening strains:
adopting a plate confronting test, placing pathogenic bacteria indicator in the center of a PDA plate, carrying out point contact on purified bacteria to be screened at two sides 2cm away from the indicator bacteria block, culturing for 4d in a constant-temperature incubator at 37 ℃, observing the growth conditions of two bacterial colonies, observing whether a bacteriostatic zone is generated, and screening out the bacterial strain with the strongest inhibition capacity on the pathogenic bacteria. FIG. 3 is a graph showing the results of the plate confrontation experiment of the strains with the highest bacteriostatic ability and pathogenic bacteria. The pathogenic bacteria indicator bacteria are fusarium oxysporum.
(2) And (3) strain identification:
extracting genome DNA of the strain, amplifying a 16S rDNA sequence, and comparing in an NCBI database, wherein the preliminary identification method comprises the following steps: extracting genome DNA of the strain by adopting a genome extraction kit, and performing amplification reaction by adopting a 16S rDNA universal primer upstream primer 27F: 5'-AGA GTT TGA TCC TGG CTC AG 3', downstream primer (1492R): the 16S rDNA sequence was amplified by 5'-TAC GGY TAC CTT GTT ACG ACT T3' PCR, and the PCR product was sent to Biotechnology engineering (Shanghai) GmbH for sequencing.
The sequencing result is as follows: the 16S rDNA sequence of Bacillus amyloliquefaciens is shown in SEQ ID NO 1.
The comparison is carried out in NCBI database, and the sequence comparison result is as follows: the homology with partial sequence of bacillus amyloliquefaciens is more than 99 percent. Its morphological characteristics on the plate (fig. 1): the colony surface is not smooth and white, the edge is irregular, and the microscopic image is as shown in FIG. 2: is in a long rod shape, and spores are formed in the center.
The preservation information of YQ1-2 obtained by screening in the invention is as follows:
the preservation name is Bacillus amyloliquefaciens, and the preservation unit is as follows: china general microbiological culture Collection center, preservation Address: xilu No.1 Hospital No. 3, Beijing, Chaoyang, on Beijing, with a deposit number: CGMCC No.20309, preservation time 2020, 07/2020.
(3) Effect of different inoculum size on growth of the strain:
inoculating into LB culture medium with the same volume at the inoculation amount of 0.5%, 1.0%, 2.0% and 4%, respectively, placing on a shaker at 30 deg.C, measuring OD every 1h with spectrophotometer600Value, measured for 48h continuously, setting 1 blank (i.e., inoculum size 0); the results are shown in FIG. 4, and the growth rate of the strain is determined to be faster, the strain enters the logarithmic growth phase 7 hours after inoculation, and the strain enters the stationary phase 24 hours after inoculation; the influence of different inoculation amounts on the growth of the strain is small, and the time of the strain entering a logarithmic growth phase and a stationary phase has no obvious difference; after 28h of culture, the thallus density is reduced when 4% of high inoculation amount is treated; the culture result shows that the strain has stronger metabolic capability and faster growth rate, and quickly occupies ecological niche in colonization;
the LB culture medium is: 5g of yeast extract, 10g of tryptone, 10g of NaCl, and pH 7.0.
(4) Effect of different pH on strain growth:
adjusting the pH of LB culture medium to 4.0, 5.0, 6.0, 7.0, 8.0, sterilizing at 121 deg.C for 30min, inoculating the strain to the culture medium at an inoculum size of 1%, placing on a shaker at 29-30 deg.C, and using at intervals of 1 hrSpectrophotometric determination of OD600Value, measured for 48h continuously, set 1 blank control; as shown in FIG. 5, pH affected the growth of the strain significantly, and at pH 4.0, the growth rate of the strain was low and no growth occurred at the end of the culture; the strain showed the highest growth rate at initial pH 6 and pH7. The log phase and stationary phase of the strains did not differ much at each pH.
(5) Effect of different NaCl concentrations on the growth of the strain:
adjusting the NaCl concentration of the LB culture medium as follows: 0.5%, 1.0%, 2.0% and 4%, inoculating the strain into culture medium at an inoculation amount of 1%, placing on a shaker at 29-30 deg.C, measuring OD every 1h by spectrophotometry600Value, measured for 48h continuously, set 1 blank control; the results are shown in fig. 6, the NaCl concentration has a greater effect on the growth of the strain, the strain grows fastest at a NaCl concentration of 1%, and is lower or higher than 1%, which has a certain inhibitory effect on the strain.
(6) Effect of different temperatures on growth of the strain (including effect on sporulation of the strain):
inoculating the strain to a culture medium with an inoculum size of 1%, and culturing at 25 deg.C, 30 deg.C (24h) -40 deg.C (30 deg.C for 24h and then 40 deg.C), 35 deg.C and 40 deg.C respectively; after 24h (entering log phase), the OD was measured by spectrophotometry600The value is obtained. The spore formation was observed with a microscope every 5 hours from 15 hours, and the spore formation rate was calculated. The results are shown in fig. 7 and fig. 8, the culture temperature has a large influence on the growth of the strain, wherein the strain grows fastest at 30 ℃ and 30 ℃ (24h) -40 ℃, and the strain can be inhibited at lower temperature or higher temperature. The temperature can also influence the spore formation of the strain, and the spore formation rate of the strain after 35 hours is far higher than that of other experimental groups under the conditions of 30 ℃ (24 hours) to 40 ℃; at a temperature of 35 ℃ it takes 50h for the spores to fully form. At 25 deg.C, no formation of spore occurs during the spore culture time, and at 30 deg.C and 50 deg.C, the maximum rate of spore formation is less than 50%.
Example 2, a microbial preparation prepared by the following method:
(1) inoculating Bacillus amyloliquefaciens YQ1-2(CGMCC No.20309) into LB culture medium according to the inoculation amount of 1%, culturing at 30 ℃ for 22-26h (preferably 24h), and continuously culturing at 40 ℃ for 18-26h (preferably 21h) to obtain liquid microbial inoculum (bacterial liquid). The spore formation rate is more than 90%, and the viable count exceeds 1x1010cfu/mL。
The LB culture medium is: 5g of yeast extract, 10g of tryptone, 10g of NaCl, and pH 7.0.
Example 3, the liquid microbial inoculum (bacterial liquid) obtained in example 2 is added into the organic fertilizer according to the addition amount of 1%, and the mixture is uniformly mixed, so that the biological organic fertilizer containing bacillus amyloliquefaciens (the number of effective viable bacteria is more than 5000 ten thousand/g) is obtained.
Example 4 field test of the bio-organic fertilizer described in example 3 for plant growth promotion and disease control
(1) Selecting melons as test crops, and respectively testing in bases of Yiwu city in Jinhua city and Jiaxing city Jiashan county in 12 months in 2019, wherein the test soil in Jiashan county is rice soil, and the weak acidity (the pH is about 6.5) of the soil is obtained; the soil to be tested in the Yiwu city is sandy loam, and the soil is acidic (the pH is about 4.56). No serious natural disasters occurred at both test points during the test.
(2) The experiment set up 3 treatments.
Treatment 1: no fertilization, CK;
and (3) treatment 2: applying an organic fertilizer without the bacillus amyloliquefaciens into soil once before melon field planting, wherein the using amount is 300 kg/mu;
and (3) treatment: the bacillus amyloliquefaciens bio-organic fertilizer is applied to soil once before melon field planting, and the using amount is 300 kg/mu.
The organic fertilizer can be produced by Sanqing agriculture development Co., Ltd, Lishui, Zhejiang province.
Jiashan test points: and (3) performing plug-seedling culture in 12 and 15 days in 2019, ploughing and fertilizing in 1 and 4 days in 2020, and transplanting seedlings in 11 days in 1 and 11 days.
And (3) artificial black test points: seeding and raising seedlings in a plug tray in 12 months and 10 days in 2019, ploughing and fertilizing in 12 months and 25 days, and transplanting seedlings in 12 months and 30 days.
The rest of the water and fertilizer management is managed according to a conventional mode.
(4) The test is a cell comparison test, 3 times of repetition are set for each treatment, 9 cells are set in total, 56 plants are arranged in each cell, and the cells are randomly arranged in groups. Regularly observing the growth vigor of the melons during the test period, measuring the height from the root base to the top of each planting plant after planting for 1 month as the height of each processed melon plant, and measuring the stem thickness of the 10 th section above the root before ripe picking of the melons (about 3 months after planting); and (3) regularly observing and recording the incidence conditions of soil-borne diseases such as sweet melon fusarium wilt, root rot and the like during the test period, counting the final incidence number of each treatment, and calculating the incidence rate according to the ratio of the incidence number to the total planting number.
(5) The effect of different fertilization treatments on melon growth is shown in table 1. As shown in the table, the biological organic fertilizer containing the bacillus amyloliquefaciens has obvious growth promoting effect on melons and can also obviously reduce the incidence rate of the melons in growth. The melon growth conditions (Jiashan test points) are shown in FIG. 9.
TABLE 1 Effect of different fertilisation treatments on melon growth
Note: the same column of different letters indicates significance of difference at a P <0.05 level
Comparative experiment, the "applying bacillus amyloliquefaciens bio-organic fertilizer of the invention" in the example 4 is changed into the bacterial liquid shown in the following table, the viable count of the organic fertilizer is kept unchanged, the rest is equal to the example 4, the comparative experiment is only carried out in the Nippon jews, and the obtained result is as follows:
TABLE 2 influence of different bacterial solutions on melon growth
Finally, it is also noted that the above-mentioned lists merely illustrate a few specific embodiments of the invention. It is obvious that the invention is not limited to the above embodiments, but that many variations are possible. All modifications which can be derived or suggested by a person skilled in the art from the disclosure of the present invention are to be considered within the scope of the invention.
Sequence listing
<110> original technology (Hangzhou) Co., Ltd
<120> growth-promoting disease-resistant bacillus amyloliquefaciens and application thereof
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<400> 1
tcaccgactt cgggtgttac aaactctcgt ggtgtgacgg gcggtgtgta caaggcccgg 60
gaacgtattc accagcggca tgctgatccg cgattactag cgattccagc ttcacgcagt 120
cgagttgcag actgcgatcc gaactgagaa cagatttgtg ggattggctt aacctcgcgg 180
tttcgctgcc ctttgttctg tccattgtag cacgtgtgta gcccaggtca taaggggcat 240
gatgatttga cgtcatcccc accttcctcc ggtttgtcac cggcagtcac cttagagtgc 300
ccaactgaat gctggcaact aagatcaagg gttgcgctcg ttgcgggact taacccaaca 360
tctcacgaca cgagctgacg acaaccatgc accacctgtc actctgcccc cgaaggggac 420
gtcctatctc taggattgtc agaggatgtc aagacctggt aaggttcttc gcgttgcttc 480
gaattaaacc acatgctcca ccgcttgtgc gggcccccgt caattccttt gagtttcagt 540
cttgcgaccg tactccccag gcggagtgct taatgcgtta gctgcagcac taaggggcgg 600
aaacccccta acacttagca ctcatcgttt acggcgtgga ctaccagggt atctaatcct 660
gttcgctccc cacgctttcg ctcctcagcg tcagttacag accagagagt cgccttcgcc 720
actggtgttc ctccacatct ctacgcattt caccgctaca cgtggaattc cactctcctc 780
ttctgcactc aagttcccca gtttccaatg accctccccg gttgagccgg gggctttcac 840
atcagactta agaaaccgcc tgcgagccct ttacgcccaa taattccgga caacgcttgc 900
cacctacgta ttaccgcggc tgctggcacg tagttagccg tggctttctg gttaggtacc 960
gtcaaggtgc cgccctattt gaacggcact tgttcttccc taacaacaga gctttacgat 1020
ccgaaaacct tcatcactca cgcggcgttg ctccgtcaga ctttcgtcca ttgcggaaga 1080
ttccctactg ctgcctcccg taggagtctg ggccgtgtct cagtcccagt gtggccgatc 1140
accctctcag gtcggctacg catcgtcgcc ttggtgagcc gttacctcac caactagcta 1200
atgcgccgcg ggtccatctg taagtggtag ccgaagccac cttttatgtc tgaaccatgc 1260
ggttcagaca accatccggt attagccccg gtttcccgga gttatcccag tcttacaggc 1320
aggttaccca cgtgttactc acccgtccgc cgctaacatc agggagcaag ctccc 1375
Claims (5)
1. Bacillus amyloliquefaciens YQ1-2, which is characterized in that: is Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) YQ1-2 with the preservation number as follows: CGMCC No. 20309.
2. The use of bacillus amyloliquefaciens YQ1-2 according to claim 1, wherein: has growth promoting effect on plants.
3. The use of bacillus amyloliquefaciens YQ1-2 according to claim 2, wherein: improve the disease resistance of plants.
4. The use of bacillus amyloliquefaciens YQ1-2 according to claim 2 or 3, wherein: the plant is a melon.
5. The use of Bacillus amyloliquefaciens YQ1-2 according to claim 4, wherein: has disease-resistant effect on the blight and root rot of sweet melons.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101948771A (en) * | 2010-08-27 | 2011-01-19 | 黑龙江省科学院微生物研究所 | Bacillus amyloliquefaciens growing in disease-preventing and growth-promoting plant and application thereof |
| CN105132336A (en) * | 2015-10-10 | 2015-12-09 | 中国热带农业科学院环境与植物保护研究所 | Bacillus amyloliquefaciens and microbial inoculum and application thereof |
| CN107245461A (en) * | 2017-05-31 | 2017-10-13 | 北方民族大学 | A kind of bacillus amyloliquefaciens B6 and its application |
| CN109006240A (en) * | 2018-07-16 | 2018-12-18 | 山东省农业科学院蔬菜花卉研究所 | A method of increase production for continuous cropping second crop soil melon crop promoting root growth |
-
2020
- 2020-12-07 CN CN202011427349.5A patent/CN112522146A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101948771A (en) * | 2010-08-27 | 2011-01-19 | 黑龙江省科学院微生物研究所 | Bacillus amyloliquefaciens growing in disease-preventing and growth-promoting plant and application thereof |
| CN105132336A (en) * | 2015-10-10 | 2015-12-09 | 中国热带农业科学院环境与植物保护研究所 | Bacillus amyloliquefaciens and microbial inoculum and application thereof |
| CN107245461A (en) * | 2017-05-31 | 2017-10-13 | 北方民族大学 | A kind of bacillus amyloliquefaciens B6 and its application |
| CN109006240A (en) * | 2018-07-16 | 2018-12-18 | 山东省农业科学院蔬菜花卉研究所 | A method of increase production for continuous cropping second crop soil melon crop promoting root growth |
Non-Patent Citations (1)
| Title |
|---|
| 李佳 等: "微生物菌剂处理重复利用基质的研究与评价", 《现代农业科技》 * |
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