CN112812995B - Paenibacillus polymyxa capable of preventing and treating root rot of salvia miltiorrhiza and application thereof - Google Patents
Paenibacillus polymyxa capable of preventing and treating root rot of salvia miltiorrhiza and application thereof Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G13/00—Protecting plants
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G17/00—Cultivation of hops, vines, fruit trees, or like trees
- A01G17/005—Cultivation methods
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G7/00—Botany in general
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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Abstract
The strain number of the paenibacillus polymyxa (Paenibacillus polymyxa) Pp2-1 is preserved in China center for type culture Collection (CCTCC for short), and the preservation address is as follows: the preservation number of the registration book is CCTCC M2020929, and the preservation date is 2020, 12 months and 21 days. The Paenibacillus polymyxa has a good inhibiting effect on root rot pathogenic bacteria of salvia miltiorrhiza, has a prevention effect on root rot diseases of salvia miltiorrhiza of more than 28%, can effectively reduce the morbidity of the root rot diseases of salvia miltiorrhiza, and has an effect superior to that of the existing common chemical pesticide. After the screened paenibacillus polymyxa is used, the chemical pesticide common problems such as drug resistance, pesticide residue, environmental pollution and the like can not be generated, and the bacillus polymyxa is healthy and environment-friendly to people and livestock and meets the requirements of people on ecological agriculture.
Description
Technical Field
The invention relates to paenibacillus polymyxa capable of preventing and treating root rot of salvia miltiorrhiza and application thereof, belonging to the technical field of microorganisms.
Background
Salvia miltiorrhiza Salvia milithioriza bge is a perennial herb of Labiatae, is taken as a medicine by drying roots and rhizomes, and is one of the first-choice medicines for treating cardiovascular and cerebrovascular diseases. In recent years, the planting of salvia has become an important measure for the poverty elimination and the countryside revivification in poverty-stricken areas in western Henan province. In order to ensure the yield of the salvia miltiorrhiza, most medicinal material growers widely use chemical fertilizers and chemical agents in the planting process of the salvia miltiorrhiza, so that the quality of the salvia miltiorrhiza is reduced to a certain extent, fertilizer damage and chemical damage can also occur under severe conditions, even chemical agent residue exceeds the standard, and meanwhile, the ecological environment of a planting area is polluted. The utilization of beneficial antagonistic bacteria for biological control and soil regulation is a research hotspot which is concerned by the current time. At present, the biological mode of microorganisms is mainly relied on to control plant diseases, including bacteria, fungi, actinomycetes and the like. The root rot of salvia miltiorrhiza is the most important disease in the production of salvia miltiorrhiza, and the yield of salvia miltiorrhiza is seriously reduced. The root rot pathogens most threatening salvia miltiorrhiza mainly include fusarium (fusarium proliferatum), fusarium solani (f.solani), fusarium oxysporum (f.oxysporum) and alternaria tenuissima (alternaria tenuissima) of fusarium genus. Biological control has been paid attention by students in the aspect of controlling root rot diseases of salvia miltiorrhiza, biocontrol strain screening of pathogenic bacteria F.solani of the root rot disease of salvia miltiorrhiza and microbial inoculum and application thereof have been widely researched, and it has been disclosed that biocontrol strains mainly comprise bacillus subtilis, bacillus amyloliquefaciens, trichoderma and the like, and have fewer effective biocontrol strains on pathogenic bacteria F.proliferum, F.oxysporum and Alternaria alternata, so that screening of biocontrol strains which are effective simultaneously on different pathogenic bacteria of the root rot disease of salvia miltiorrhiza has important theoretical and practical significance.
Disclosure of Invention
Aiming at the defects of the prior art, the invention aims to provide Paenibacillus polymyxa capable of preventing and treating root rot of salvia miltiorrhiza and application thereof, and a screening method and application of the strain.
The technical scheme adopted by the invention for solving the technical problems is as follows:
the strain is numbered Paenibacillus polymyxa (Paenibacillus polymyx) Pp2-1, is preserved in China center for type culture Collection (CCTCC for short), and has the preservation address: the preservation number of the Shandong type culture Collection of Wuhan university, eight Hongshan areas, Hubei province, is CCTCC M2020929, and the preservation date is 2020, 12 months and 21 days.
The Paenibacillus polymyxa capable of preventing and treating root rot of red-rooted salvia root is prepared by the following screening method: collecting salvia miltiorrhiza rhizosphere soil in a salvia miltiorrhiza field, weighing 1g of soil, adding 10ml of sterile water, fully oscillating to obtain a soil suspension, shaking the soil suspension uniformly, diluting, coating 0.2ml of diluent on an LB solid plate, culturing, separating and purifying after bacterial colonies grow out, screening out a strain with an inhibiting effect on salvia miltiorrhiza root rot by using a plate antagonism test, and identifying to obtain the paenibacillus polymyxa Pp 2-1.
The Paenibacillus polymyxa capable of preventing and treating root rot of salvia miltiorrhiza bunge is applied to prevention and treatment of root rot of salvia miltiorrhiza bunge.
The paenibacillus polymyxa capable of preventing and treating root rot of salvia miltiorrhiza is applied to preparation of a microbial inoculum for preventing and treating root rot of salvia miltiorrhiza.
The preparation method of the microbial inoculum comprises the following steps: the method comprises the steps of inoculating paenibacillus polymyxa Pp2-1 into an LB culture medium, carrying out shake culture for 12 hours at 28 ℃ and 180rpm to prepare seed bacteria, inoculating the seed bacteria into the LB culture medium in an inoculum size of 2%, and fermenting for 48 hours at 28 ℃ and 180rpm to form fermentation liquor, namely the microbial inoculum.
The concentration of the live bacteria of the paenibacillus polymyxa Pp2-1 in the fermentation liquor is 10 7 ～10 8 CFU/mL。
The LB culture medium is as follows: 10g of tryptone, 5g of yeast extract and 10g of sodium chloride, adding distilled water to a constant volume of 1000mL, uniformly mixing, subpackaging in triangular flasks, and sterilizing at 121 ℃ for 30 min.
The using method of the microbial inoculum comprises the following steps: the microbial inoculum is diluted by 10 times by sterile water to ensure that the viable bacteria concentration of the diluted solution is 10 6 ～10 7 CFU/ml, irrigating root around root of Saviae Miltiorrhizae radix; the root irrigation is carried out 1 time every 7 days, and the root irrigation is carried out 2 times.
The invention has the beneficial effects that: the invention aims to prevent and treat root rot of salvia miltiorrhiza by using microorganisms, so Pp2-1 capable of preventing and treating root rot of salvia miltiorrhiza is separated from rhizosphere soil of salvia miltiorrhiza through separation identification, plate antagonism and disease prevention potting experiments, and is identified as paenibacillus polymyxa. Tests show that the paenibacillus polymyxa has a good inhibiting effect on root rot pathogenic bacteria of salvia miltiorrhiza, the control effect on the root rot diseases of salvia miltiorrhiza can reach more than 28%, the morbidity of the root rot diseases of salvia miltiorrhiza can be effectively reduced, and the effect is superior to that of the existing common chemical pesticide. The Paenibacillus polymyxa screened by the method does not generate the common problems of chemical pesticides such as drug resistance, pesticide residue, environmental pollution and the like after being used, is healthy and environment-friendly to people and livestock, and meets the requirements of people on ecological agriculture.
Drawings
FIG. 1 is a graph comparing the results of plate antagonism tests of the strain Pp2-1 to Fusarium proliferatum, Fusarium solani, Fusarium oxysporum and Alternaria tenuissima.
FIG. 2 is a colony morphology of the strain Pp 2-1.
FIG. 3 is a comparative graph of the result of the tobacco leaf allergy test by the strain Pp2-1, wherein the left side of the graph is the strain Pp2-1, and the right side of the graph is a positive control.
Detailed Description
The present invention will be further described with reference to the following examples and the accompanying drawings.
Separation of biocontrol bacteria
Weighing 1g of soil, adding 10ml of sterile water, fully oscillating to obtain a soil suspension, shaking the soil suspension uniformly, diluting, coating 200 mu L of diluent on an LB solid plate, culturing until bacterial colonies grow out, selecting bacterial colonies with different sizes, colors or forms, streaking, separating and purifying on the surface of an LB solid culture medium to obtain 1 strain, which is named as Pp 2-1.
(II) screening of biocontrol bacteria (Flat plate antagonism test)
Screening biocontrol bacteria by a plate confronting method, respectively inoculating fusarium, fusarium solani, fusarium oxysporum and alternaria tenuissima from a root rot pathogen layer of salvia miltiorrhiza on a PDA culture medium, culturing for 5d at 28 ℃, preparing cultured root rot pathogens of salvia miltiorrhiza into bacterial cakes by using a perforator with the sterilized diameter of 8mm, connecting the bacterial cakes to one side of a new PDA plate (the diameter of 90mm), sterilizing a paper disc ring with the diameter of 8mm, placing the paper disc ring in the other side of the PDA culture medium, and ensuring that the biocontrol bacteria and the paper disc ring are positioned on the same straight line during inoculation. Sealing with sealing film, culturing at 28 deg.C, recording the size of the antibacterial band when the control pathogenic bacteria grow to the paper disc, and measuring the colony radius of the corresponding pathogenic bacteria on the plate. The contrast is the culture of Fusarium, Fusarium solani, Fusarium oxysporum and Alternaria tenuissima on the root rot pathogenic bacteria layer of the salvia miltiorrhiza and the culture of the bacteria on a PDA by inoculating a sterile water paper disc, the bacterial colony treatment is repeated for at least 3 times, and the results of opposite culture are checked after 5 days, and are shown in the table 1 and the figure 1, and Pp2-1 has effects on 4 pathogenic bacteria.
Table 1: bacteriostatic effect of paenibacillus polymyxa (Pp2-1) on root rot pathogenic bacteria of salvia miltiorrhiza
(III) identification of biocontrol bacteria
And (3) performing morphological identification on the screened strains with good antagonistic effect, and determining the strains as the B.leisiensis through morphological, physiological and biochemical analysis (see table 2) and molecular biological identification. The Bacillus belgii forms milky white colonies on the culture medium, which are opaque, slightly convex in the middle and irregular in the edge (see FIG. 2).
Table 2: physiological and biochemical characteristics of Pp2-1
Note: "-" is negative and "+" is positive.
(IV) sequencing of biocontrol bacteria
Sequencing: the strain Pp2-1 was sequenced, and the result was identified as Paenibacillus polymyxa (Paenibacillus polymyxa).
The 16SrDNA gene sequencing result is as follows:
SEQUENCE LISTING
<110> institute of plant protection, institute of agricultural sciences, Henan province
<120> Paenibacillus polymyxa capable of preventing and treating root rot of salvia miltiorrhiza and application thereof
<130>2021.1.5
<160>1
<170>PatentIn version 3.5
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(V) preparation of microbial inoculum
Inoculating paenibacillus polymyxa Pp2-1 into an LB culture medium, carrying out shake culture for 12h at 28 ℃ and 180rpm to prepare seed bacteria, inoculating the seed bacteria into the LB culture medium in an inoculum size of 2%, and fermenting for 48h at 28 ℃ and 180rpm to form fermentation liquor, namely the microbial inoculum. The concentration of the live bacteria of the paenibacillus polymyxa Pp2-1 in the fermentation liquor is 10 7 ～10 8 CFU/mL。
(VI) test of tobacco allergy of microbial inoculum
The tobacco lower epidermis was inoculated subcutaneously with 10mM MgSO 4 As a negative control, the wild tobacco fire was used as a positive control (YH). Under the conditions of 24 ℃ and 60% -80% high humidity, allergic necrotic spots appear after 24-48h as positive reaction, and yellow spots appear after 3d as negative reaction. As a result, as shown in FIG. 3, the anaphylaxis test of the Paenibacillus polymyxa Pp2-1 is a negative reaction, so that the Paenibacillus polymyxa Pp2-1 is nonpathogenic and can be used as a biocontrol bacterium for controlling the root rot of salvia miltiorrhiza.
(VII) disease prevention experiment of biocontrol bacteria
Germinating Saviae Miltiorrhizae radix seed on a tray filled with sterilized soil, transferring into nutrition pot, culturing for 30 days, inoculating biocontrol bacteria by root irrigation method, wetting soil with small amount of clear water before root irrigation, inoculating Paenibacillus polymyxa Pp2-1 fermentation broth (viable bacteria concentration is 10) at rhizosphere 7 ～10 8 CFU/mL), diluting the fermentation broth 10 times with sterile water, 10mL for each plant, and inoculating 10mL again at intervals of 7 d; the control group is inoculated with an equal amount of LB culture medium (CK) by the same method, in addition, a medicament positive control group is arranged, the control medicament is 1000-time diluent of 50 percent carbendazim wettable powder, each salvia miltiorrhiza seedling is 10mL, and after 7 days, the roots are irrigated for 1 time and 2 times in total. After the root is irrigated for 3d for the last time in each group, the fusarium solani which is the pathogen of the root rot of salvia miltiorrhiza is inoculated. The inoculation method comprises the following steps: irrigating root and inoculating, taking 10mL root rot pathogen spore suspension (concentration is 1X 10) 6 root/mL) of Salvia miltiorrhiza seedling, watering for 1 time in 3d, culturing in 28 deg.C incubator, and periodically observing disease of Salvia miltiorrhizaThe situation is. And after culturing for 30 days, investigating and calculating the morbidity, disease index and prevention and treatment effect. The grading standard of the root rot disease of salvia miltiorrhiza is carried out according to related documents, and the result is shown in table 3. The result shows that the control effect of the paenibacillus polymyxa Pp2-1 on the root rot of salvia miltiorrhiza is 28.1%.
TABLE 3 Paenibacillus polymyxa Pp2-1 disease prevention effect on salvia miltiorrhiza root rot pot culture
The result shows that the control effect of the paenibacillus polymyxa Pp2-1 on the root rot of salvia miltiorrhiza is 28.1%. The effect is obvious. The foregoing is only a preferred embodiment of the invention and various modifications and changes will occur to those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Sequence listing
<110> institute of plant protection of academy of agricultural sciences of Henan province
<120> Paenibacillus polymyxa capable of preventing and treating root rot of salvia miltiorrhiza and application thereof
<141> 2021-01-07
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agcccagaga gtcgccttcg ccactggtgt tcctccacat ctctacgcat ttcaccgcta 840
cacgtggaat tccactctcc tcttctgcac tcaagctctc cagtttccag tgcgacccga 900
agttgagcct cgggattaaa caccagactt aaagagccgc ctgcgcgcgc tttacgccca 960
ataattccgg acaacgcttg ccccctacgt attaccgcgg ctgctggcac gtagttagcc 1020
ggggctttct tctcaggtac cgtcactctt atagcagtta ctctacaaga cgttcttccc 1080
tggcaacaga gctttacgat ccgaaaacct tcatcactca cgcggcgttg ctccgtcagg 1140
ctttcgccca ttgcggaaga ttccctactg ctgcctcccg taggagtctg ggccgtgtct 1200
cagtcccagt gtggccgatc accctctcag gtcggctacg catcgtcgcc ttggtaggcc 1260
tttaccccac caacaagcta atgcgccgca ggcccatcca caagtgacag attactccgt 1320
ctttcctcct tctcccatgc aggaaaagga tgtatcgggt attagctacc gtttccggta 1380
gttatccctg tcttgtgggc aggttgccta cgtgttactc acccgtccgc cgctaggttg 1440
tttagaagca agcttctaaa caacctcgct cgacttgcat gtattaggca cgccgccagc 1500
gttcgtcctg agccaggatc aaactct 1527
Claims (6)
1. A Paenibacillus polymyxa for preventing and treating root rot of red sage root is disclosed, which is numbered as Paenibacillus polymyxa (A), (B), (C), (Paenibacillus polymyx) Pp2-1, which has been preserved in China center for type culture Collection (CCTCC for short), the preservation address is: the preservation number of the registration book is CCTCC M2020929, and the preservation date is 2020, 12 months and 21 days.
2. The use of paenibacillus polymyxa according to claim 1 for the control of root rot of salvia miltiorrhiza.
3. The application of the paenibacillus polymyxa according to claim 1 in preparing a microbial inoculum for preventing and treating root rot of salvia miltiorrhiza.
4. The application of claim 3, wherein the preparation method of the microbial inoculum comprises the following steps: inoculating paenibacillus polymyxa Pp2-1 into an LB culture medium, carrying out shake culture for 12h at 28 ℃ and 180rpm to prepare seed bacteria, inoculating the seed bacteria into the LB culture medium in an inoculum size of 2%, and fermenting for 48h at 28 ℃ and 180rpm to form fermentation liquor, namely the microbial inoculum.
5. The method for preparing the bacterial agent according to claim 4, wherein the concentration of the viable bacteria of Paenibacillus polymyxa Pp2-1 in the bacterial agent is 10 7 ～10 8 CFU/mL。
6. The use of claim 3, wherein the microbial inoculum is used by a method comprising: the microbial inoculum is diluted by 10 times by sterile water to ensure that the viable bacteria concentration of the diluted solution is 10 6 ～10 7 CFU/ml, irrigating root around root of Saviae Miltiorrhizae radix; the root irrigation is carried out 1 time every 7 days, and the root irrigation is carried out 2 times.
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Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004050861A1 (en) * | 2002-12-04 | 2004-06-17 | Shanghai Zeyuan Marine Biotechnology Ltd. | The microbial preparation & method for preventing and curing the bacterial wilt the plant and its use |
| WO2015100431A2 (en) * | 2013-12-24 | 2015-07-02 | Symbiota, Inc. | Plants containing beneficial endophytes |
| CN105132324A (en) * | 2015-09-07 | 2015-12-09 | 哈尔滨师范大学 | Wheat rhizosphere paenibacillus polymyxa WXD 6-4 and application thereof |
| CN105578884A (en) * | 2013-07-24 | 2016-05-11 | 拜耳作物科学股份公司 | Binary fungicidal composition |
| CN105733990A (en) * | 2016-03-30 | 2016-07-06 | 青岛蔚蓝生物集团有限公司 | Paenibacillus polymyxa and application thereof in prevention and control of plant diseases |
| CN107619334A (en) * | 2016-07-21 | 2018-01-23 | 济南航晨生物科技有限公司 | A kind of special ocean type disease-resistant compound microbial fertilizer of the red sage root |
| CN107937326A (en) * | 2018-01-17 | 2018-04-20 | 鲁东大学 | A kind of Paenibacillus polymyxa and its application for being used to prevent American ginseng root maize ear rot |
| CN111748509A (en) * | 2020-07-03 | 2020-10-09 | 浙江工商大学 | Microbial purifying preparation for seawater culture tail water, and preparation method and application thereof |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2016187703A1 (en) * | 2015-05-22 | 2016-12-01 | Institut National De La Recherche Scientifique | Bacterial and fungal metabolites possessing anti-microbial activity against xanthomonas species, compositions, methods, kits and uses relating to same |
-
2021
- 2021-01-07 CN CN202110019636.0A patent/CN112812995B/en not_active Expired - Fee Related
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004050861A1 (en) * | 2002-12-04 | 2004-06-17 | Shanghai Zeyuan Marine Biotechnology Ltd. | The microbial preparation & method for preventing and curing the bacterial wilt the plant and its use |
| CN105578884A (en) * | 2013-07-24 | 2016-05-11 | 拜耳作物科学股份公司 | Binary fungicidal composition |
| WO2015100431A2 (en) * | 2013-12-24 | 2015-07-02 | Symbiota, Inc. | Plants containing beneficial endophytes |
| CN105132324A (en) * | 2015-09-07 | 2015-12-09 | 哈尔滨师范大学 | Wheat rhizosphere paenibacillus polymyxa WXD 6-4 and application thereof |
| CN105733990A (en) * | 2016-03-30 | 2016-07-06 | 青岛蔚蓝生物集团有限公司 | Paenibacillus polymyxa and application thereof in prevention and control of plant diseases |
| CN107619334A (en) * | 2016-07-21 | 2018-01-23 | 济南航晨生物科技有限公司 | A kind of special ocean type disease-resistant compound microbial fertilizer of the red sage root |
| CN107937326A (en) * | 2018-01-17 | 2018-04-20 | 鲁东大学 | A kind of Paenibacillus polymyxa and its application for being used to prevent American ginseng root maize ear rot |
| CN111748509A (en) * | 2020-07-03 | 2020-10-09 | 浙江工商大学 | Microbial purifying preparation for seawater culture tail water, and preparation method and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| Biocontrol of Fusarium Crown and Root Rot and Promotion of Growth of Tomato by Paenibacillus Strains Isolated from Soil;Sheng Jun Xu et al.;《Mycobiology》;20140630;第42卷(第2期);第158-166页 * |
| 一株燕麦根腐病生防菌的鉴定及其防治效果;张建强等;《草业科学》;20211231;第 38 卷(第 11 期);第2266-2273页 * |
| 抗香蕉枯萎病甲基营养型芽孢杆菌的鉴定及定殖;周登博等;《中国农学通报》;20171231;第33卷(第36期);第145-151页 * |
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