CN116790417A - Pseudomonas capable of inhibiting pathogenic bacteria of various plants and having growth promoting effect on various crops, screening method and application - Google Patents
Pseudomonas capable of inhibiting pathogenic bacteria of various plants and having growth promoting effect on various crops, screening method and application Download PDFInfo
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- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
- A01N63/27—Pseudomonas
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Abstract
The invention provides pseudomonas capable of inhibiting pathogenic bacteria of various plants and having growth promoting effect on various crops, a screening method and application thereof. The Pseudomonas is Pseudomonas A34-9 (Pseudomonas sp.A34-9) which is deposited in China center for type culture Collection, with accession number: cctccc NO: m2022397, date of preservation: 2022, 04, 08. The pseudomonas has inhibition effect on various fusarium, phytophthora and rhizoctonia solani, wherein the prevention effect on Fusarium solani is 59.69% and the prevention effect on phytophthora is 75%. The Pseudomonas can inhibit growth of various plant pathogenic fungi, and promote yield improvement of Saviae Miltiorrhizae radix, rhizoma Dioscoreae, atractylodis rhizoma, flos Lonicerae, wheat and peanut.
Description
Technical Field
The invention relates to pseudomonas capable of inhibiting pathogenic bacteria of various plants and having growth promoting effect on various crops, a screening method and application thereof, and belongs to the technical field of microorganisms.
Background
Along with the requirements of sustainable development strategy of modern agriculture, the biological control means not only meets the strategic aim, but also can effectively promote the development of the agriculture industry, and at present, biological control has been widely paid attention to the aspect of soil-borne disease control, wherein root rot, wilt and black shank are common soil-borne diseases, and various fusarium, rhizoctonia solani, phytophthora and the like are main pathogens causing diseases of various crops such as root rot, fusarium wilt, rhizoctonia solani, black shank and the like. The promotion effect of biocontrol bacteria on crop growth is also receiving a great deal of attention while controlling disease occurrence and recovering crop yield and quality loss. The use of biocontrol bacteria to replace chemical pesticides and fertilizers is becoming a research hotspot of modern agricultural technology. The biocontrol bacteria for promoting the crop production to control the occurrence of diseases can promote the growth and development of plants by means of phosphate dissolving, potassium dissolving, nitrogen fixing, production of plant growth hormone such as indoleacetic acid IAA and the like. In recent years, researches on the growth promoting effect and IAA-producing bacteria on various crops have been recently reported, and therefore, screening of strains capable of producing IAA and promoting the growth of crops has important theoretical and practical significance.
Disclosure of Invention
Aiming at the defects of the prior art, the invention aims to provide pseudomonas capable of inhibiting pathogenic bacteria of various plants and having growth promoting effect on various crops, a screening method and application. The Pseudomonas can inhibit growth of various plant pathogenic fungi, and promote yield improvement of Saviae Miltiorrhizae radix, rhizoma Dioscoreae, atractylodis rhizoma, flos Lonicerae, wheat and peanut.
In order to achieve the above purpose, the technical scheme adopted by the invention is as follows:
pseudomonas capable of inhibiting plant pathogenic bacteria and having growth promoting effect, which is Pseudomonas A34-9 (Pseudomonas sp.A34-9) and is deposited in China center for type culture Collection, accession number: cctccc NO: m2022397, date of preservation: 2022, 04, 08, deposit address: university of martial arts in chinese.
The plant pathogenic bacteria include: fusarium, rhizoctonia solani, and phytophthora.
The fusarium comprises: fusarium asiaticum, fusarium equisetum, fusarium graminearum, fusarium pseudograminearum, fusarium layering, fusarium putrescens, fusarium oxysporum.
Plants which inhibit plant pathogenic bacteria and have a growth promoting effect include: radix Salviae Miltiorrhizae, rhizoma Dioscoreae, rhizoma Atractylodis Macrocephalae, flos Lonicerae, semen Tritici Aestivi, and semen arachidis hypogaeae.
According to the screening method of the pseudomonas, rhizosphere soil for planting tobacco on the Alshan of inner Mongolia is collected, the soil is added into sterile water to be fully oscillated to obtain a soil suspension, then the soil suspension is subjected to multiple dilution, the dilution is coated on KB solid culture medium, the culture is inverted and carried out for 36 hours in a constant temperature incubator at 28 ℃, and after bacterial colonies grow out, the bacterial strains are separated and purified, and the bacterial strains are screened.
The preparation method of the fermentation liquor when the pseudomonas is applied to the plant growth promotion comprises the following steps: inoculating pseudomonas A34-9 into KB culture medium, and carrying out shaking culture at 28 ℃ and 180rpm for 16 hours to prepare seed bacteria; inoculating seed bacteria into KB culture medium with an inoculum size of 3% (v/v), and fermenting at 28 ℃ and 180rpm for 48 hours to form fermentation liquor; the concentration of the pseudomonas viable bacteria in the fermentation liquor is 10 9 ~10 10 CFU/mL。
KB medium included: 20g of peptone, 10mL of glycerol, 1.5g of dipotassium hydrogen phosphate, 1.5g of magnesium sulfate heptahydrate and 0g or 20g of agar are added with distilled water to a volume of 1000mL, and after uniform mixing, the mixture is packaged in triangular flasks and sterilized at 121 ℃ for 30min.
The pseudomonas is applied to inhibiting plant pathogenic bacteria.
The application of the pseudomonas in plant growth promotion.
The pseudomonas is applied to the prevention and treatment of crop soil-borne root diseases.
The invention has the beneficial effects that:
the invention screens out biocontrol bacteria P34-9 which can simultaneously prevent and treat various soil-borne fungus diseases and improve crop yield from soil on Alshan for the first time through experiments of plate counter bacteriostasis, disease prevention and field growth promotion, and the strain is identified as Pseudomonas sp.A34-9. Experiments show that the pseudomonas has inhibition effect on various fusarium, phytophthora and rhizoctonia solani, wherein the prevention effect on Fusarium solani is 59.69% and the prevention effect on phytophthora is 75%.
The pseudomonas of the invention can biologically prevent and treat soil-borne diseases caused by plant pathogenic fungi such as various fusarium (fusarium oxysporum, fusarium putrescens, fusarium layering, fusarium pseudograminearum, fusarium graminearum, fusarium equisetum), rhizoctonia, phytophthora and the like, and can improve the yield of crops such as red sage root, chinese yam, white atractylodes rhizome, honeysuckle, wheat, peanut and the like.
The pseudomonas screened by the invention has good use effect, can not generate common problems of chemical pesticides such as drug resistance, pesticide residue, environmental pollution and the like after being used, is healthy to people and livestock, is environment-friendly, and meets the requirements of people on ecological agriculture.
Drawings
FIG. 1 is a comparison of growth promotion of roots and stems of Salvia Miltiorrhiza in field application for strain A34-9 of example 6.
In the figure, CK: control treatment, 3: strain A34-9 treatment.
FIG. 2 is a comparison of yam rhizome growth promotion in field applications for strain A34-9 of example 8.
FIG. 3 shows the effect of strain A34-9 on peanut field growth in example 12.
Detailed Description
The following describes the embodiments of the present invention in further detail with reference to examples.
Experimental materials:
KB medium: 20g of peptone, 10ml of glycerol, 1.5g of dipotassium hydrogen phosphate, 1.5g of magnesium sulfate heptahydrate and 20g of agar (added when only a solid culture medium is used), adding distilled water to a volume of 1000ml, packaging into triangular flasks after uniformly mixing, and sterilizing for 30min at 121 ℃.
PDA medium: 200g of potato (cut into small pieces with a side length of about 1cm, put into 800ml of distilled water and boiled for 15min, filtered by double-layer gauze to obtain filtrate), 20g of glucose, 20g of agar, adding distilled water to 1000ml, and sterilizing at 121 ℃ for 30min.
Pathogenic bacteria and other raw materials and reagents used in the invention are all purchased in the market.
Example 1 isolation of biocontrol bacteria
Collecting rhizosphere soil for planting tobacco on the Alshan of inner Mongolia, weighing 1g of soil, adding 10ml of sterile water, fully oscillating to obtain a soil suspension, shaking the soil suspension uniformly, performing double dilution, coating 0.1ml of dilution on a KB solid culture medium, culturing for 36h in an incubator at a constant temperature of 28 ℃ in an inverted manner until colonies grow out, picking colonies with different sizes, colors or forms, scribing, separating and purifying on the surface of the KB solid culture medium to obtain 1 strain, and naming the strain as P34-9.
EXAMPLE 2 inhibition of various fungal pathogens by biocontrol bacteria
Screening biocontrol bacteria by using a plate counter method: inoculating various fungal pathogens (see Table 1 in detail) on PDA culture medium, culturing at 28deg.C for 5-10 d, and punching bacterial colony with sterilized puncher into round mycelium blocks with diameter of 5 mm. The pathogenic bacteria block and the separated strain (P34-9) are inoculated onto a PDA culture medium, the distance between the pathogenic bacteria block and the separated strain is 20mm, and meanwhile, the pathogenic bacteria block is singly inoculated to serve as a control. After 3 replicates of each treatment and after the plates were filled with the control fungi, the opposite culture results were examined and shown in Table 1, with P34-9 having a certain bacteriostatic effect against a number of pathogenic bacteria (Table 1).
TABLE 1 bacteriostasis results of 1 Strain isolated from Alshan soil against various fungal pathogens
Determination of bacteriostatic activity of fermentation filtrate of isolated strain: taking bacterial strain P34-9 cultured for 24 hours, inoculating 100ml KB culture solution, shaking and fermenting for 2 days at 28 ℃ and 180rpm, centrifuging, reserving filtrate, filtering for 2 times by using a microporous filter membrane (preserving at 4 ℃), preparing a PDA plate containing 300ml/l of filtrate, inoculating 1 pathogenic bacteria block of 5mm in the center of each plate, culturing at constant temperature at 28 ℃, and observing the result after 5 days.
The result shows that the strain P34-9 has good inhibition effect on phytophthora, fusarium putrescens and Fusarium oxysporum.
Example 3 identification of biocontrol bacteria
Strain morphology and physiological biochemistry: bacterial strain P34-9 was gram-negative, and was grown in KB medium at 28℃for 48h with circular protrusions in colony morphology, smooth and moist surface, milky to yellowish, translucent, picking up the colony sticky, with diffusible fluorescent pigment (visible only on UV 2). Can grow at 5 ℃ and can not grow at 41 ℃. Arginine ditolylase, oxidase and gelatin were positive for liquefaction, nitrate reduction and denitrification tests, methyl red and VP tests were negative, and starch could not be hydrolyzed.
Sequencing: the genome sequencing is carried out on the strain P34-9, the identification result shows that the nucleic acid sequence has higher scores with Pseudomonas fluorescens (Pseudomonas fluorescens strain ATCC 13525) and Pseudomonas RU47 (P.sp.RU47), and the similarity between the strain and Pseudomonas RU47 (P.sp.RU47) is 91 percent at the highest according to the ANI analysis result, so that the strain is comprehensively identified as Pseudomonas sp.A34-9A 34-9. A34-9 was subjected to gene sequencing under the GenBank accession number CP119967.
Pseudomonas sp.A34-9, deposited with the China center for type culture Collection, accession number: cctccc NO: m2022397, date of preservation: 2022, 04, 08, deposit address: university of martial arts in chinese.
Example 4 biocontrol bacteria allergy test to tobacco leaf
Pseudomonas A34-9 was cultured at 28℃for 24 hours with 10mM MgSO 4 Formulated as 10 7 ~10 8 CFU/ml(OD 600 Inoculating common tobacco leaf by injection in bacterial liquid of 0.3-0.5), inoculating common tobacco leaf by injection at 24 deg.C under 60-80% high humidity for 24-48 hr to obtain positive reaction of allergic necrotic spot, and inoculating tobacco leaf by injection after 3d to obtain negative reaction of transparent macula lutea, inoculating tobacco leaf by injection in the presence of 10mM MgSO 4 Injection inoculation as negative control, at 10 7 ~10 8 CFU/ml tobacco wildfire pathogen injection was inoculated as a positive control.
As a result, it was found that the allergy test of Pseudomonas A34-9 was negative, and thus Pseudomonas A34-9 was nonpathogenic and could be used as a potential biocontrol bacterium.
Example 5 potted plant growth-promoting test of biocontrol bacteria on Salvia Miltiorrhiza
The preparation method of the pseudomonas A34-9 fermentation broth comprises the following steps: inoculating Pseudomonas A34-9 into KB culture medium, shake culturing at 28deg.C and 180rpm for 16 hr to obtain seed bacteria, inoculating 3% (v/v) seed bacteria into KB culture medium, fermenting at 28deg.C and 180rpm for 48 hr to obtain fermentation broth; the concentration of the pseudomonas viable bacteria in the fermentation liquor is 10 9 ~10 10 CFU/ml。
When in use, the fermentation liquid is diluted by 10 times of sterile water, and the viable bacteria concentration of the diluted liquid is 10 8 ~10 9 CFU/ml。
3-4 red sage seedlings with consistent growth vigor are selected and transplanted into flowerpots with sterilizing soil and matrix in the volume ratio of 3:1, and 1 plant is planted in each flowerpot. Inoculating pseudomonas A34-9 by root irrigation: transplanting the seedlings for 3-5 days, wetting the soil with a small amount of clean water, and then using a pipette tip to transfer the pseudomonas A34-9 fermentation liquor (10 8 CFU/ml) into plant rhizosphere, 10 ml/plant, then 7d root irrigation 1 time, 2 times total root irrigation; the control group was inoculated with an equivalent amount of KB medium (CK) in the same manner. After the biocontrol bacteria are inoculated, water is poured every 3 days, and the mixture is placed in a greenhouse at 25 ℃ for culture for 30 days, and the growth condition of the red sage root seedlings is investigated.
The result shows that the pseudomonas A34-9 has good promotion effect on the growth of the red sage root and can effectively increase the fresh weight of the red sage root (Table 2).
Table 2 promotion of growth of potted Salvia Miltiorrhiza Miq by Pseudomonas A34-9
Treatment of | Main root number (root) | Number of side roots (root) | Root weight (gram) | Root fresh weight increase yield (%) |
A34-9 | 1 | 4.13 | 2.31 | 35.09 |
CK | 1 | 4.10 | 1.71 | - |
Example 6 field growth-promoting test of biocontrol bacteria on Salvia Miltiorrhiza
The preparation method of the Pseudomonas A34-9 fermentation broth is the same as in example 5.
And (3) applying the fermentation liquor of the biocontrol bacterium pseudomonas A34-9 in the growing period of root parts of the red sage root from the bottom of June to the bottom of 7 months in the annual red sage root field with consistent growth vigor. Inoculating biocontrol bacteria by stem base spraying method, and applying 3L of Pseudomonas A34-9 fermentation liquid (living bacteria concentration is 2×10) per mu 9 CFU/liter), the inoculation was repeated 2 times at intervals of 30d, and the control group was inoculated with the same amount of KB medium (CK) by the same method. The field cultivation conditions and the water and fertilizer management are consistent, and no chemical pesticide is used any more. And (3) during the red sage harvesting period, carefully digging out the whole plant, washing off soil at the root, and measuring and recording the data of the root number, yield and the like of the red sage in the treatment and control area.
The results show that the pseudomonas A34-9 can promote root growth, root growth and root growth of the red sage root and yield of the red sage root, and the pseudomonas A34-9 has good growth promoting effect (shown in figures 1 and 3).
TABLE 3 growth promoting effect of Pseudomonas A34-9 on Salvia Miltiorrhiza
Example 7 test of biocontrol bacteria on rooting and sprouting and growth promotion of Dioscorea opposita Stem
The preparation method of the Pseudomonas A34-9 fermentation broth is the same as in example 5.
Uniformly selecting about 8cm of yam stem segments, and performing a germination and growth promotion test on the yam stem segments of the biocontrol strain Pseudomonas A34-9. The soaking method is adopted: soaking rhizoma Dioscoreae stem in Pseudomonas A34-9 fermentation broth (active bacteria concentration is 2×10) 8 CFU/liter), naturally airing after soaking, transplanting to a greenhouse sand bed for budding culture to soak KB cultureThe medium was used as a control, and a fresh water control (fresh water soaking) and a blank control (without any treatment) were set up for rooting and sprouting test.
The result shows that the Chinese yam stem segment is soaked in pseudomonas A34-9, and treated for 2 hours, the Chinese yam fibrous root length and fibrous root quantity can be obviously promoted, the growth of the overground stem is promoted, and the germination rate of the Chinese yam stem segment is improved (table 4). As can be seen from Table 4, soaking the yam stem with Pseudomonas A34-9 can effectively increase the germination rate of the yam stem and the length of the fibrous root and the length of the overground stem.
Table 4 rooting and sprouting action of Pseudomonas A34-9 on Chinese yam stem
Example 8 field growth-promoting test of biocontrol bacteria on Dioscorea opposita
The preparation method of the Pseudomonas A34-9 fermentation broth is the same as in example 5.
And (3) applying the fermentation liquor of the biocontrol pseudomonas A34-9 in the field of the Chinese yam with consistent growth vigor in the period from the bottom of June to the bottom of 7 months of underground stems of the Chinese yam. Inoculating biocontrol bacteria by stem base spraying method, and applying 2-3L of Pseudomonas A34-9 fermentation liquid (living bacteria concentration is 2×10) per mu 9 CFU/liter), the inoculation was repeated 2 times at intervals of 30d, and the control group was inoculated with the same amount of KB medium (CK) by the same method. The field cultivation conditions and the water and fertilizer management are consistent, and no chemical pesticide is used. And (3) during the harvesting period of the underground stems of the Chinese yam, carefully digging out the whole plant, washing off root soil, measuring and recording the length of the underground stems of the Chinese yam in the treatment and control area, and carrying out cell yield.
The results show that the pseudomonas A34-9 can promote the underground stem length and the single plant yield of the Chinese yam (figure 2 and table 5).
TABLE 5 growth promoting effect of Pseudomonas A34-9 on Dioscorea Panthaica
Example 9 field growth-promoting test of biocontrol bacteria on Atractylodis rhizoma
The preparation method of the Pseudomonas A34-9 fermentation broth is the same as in example 5.
And (3) applying the fermentation liquor of the biocontrol bacterium pseudomonas A34-9 in the period of vigorous growth of underground root stems of the bighead atractylodes rhizome at the bottom of June to the bottom of 7 months in the bighead atractylodes rhizome field with consistent growth vigor. Inoculating biocontrol bacteria by stem base spraying method, and respectively applying 1 liter, 2 liters and 3 liters of Pseudomonas A34-9 fermentation liquor (living bacteria concentration is 2×10) per mu land 9 CFU/liter), the inoculation was repeated 2 times at intervals of 30d, and the control group was inoculated with the same amount of KB medium (CK) by the same method. The field cultivation conditions and the water and fertilizer management are consistent, and no chemical pesticide is used any more. And (3) during the harvesting period of the underground stems of the bighead atractylodes rhizome, carefully digging out the whole plant, washing off root soil, and measuring and recording the lengths of the underground stems of the treatment and control areas in the district.
The results showed that Pseudomonas A34-9 had a significant promoting effect on the root stems of the white atractylodes rhizome (Table 6). The strain height of bighead atractylodes rhizome and the number of seedlings grown in a cell can be effectively improved by 1 liter to 2 liters per mu of pseudomonas A34-9 microbial inoculum, and the yield of the cell is increased by more than 30%.
TABLE 6 growth promoting effect of Pseudomonas A34-9 on Da Tian Baishu
Example 10 field growth-promoting test of biocontrol bacteria on honeysuckle
The preparation method of the Pseudomonas A34-9 fermentation broth is the same as in example 5.
The fermentation liquor of the biocontrol pseudomonas A34-9 is applied to the field of honeysuckle with consistent growth vigor in the bud germination period and the rapid growth period of the stem and leaf of the honeysuckle in early spring. Inoculating biocontrol bacteria by root irrigation or spraying, and applying 2L, 3L and 5L of Pseudomonas A34-9 fermentation liquid (living bacteria concentration is 2×10) respectively 9 CFU/liter), the inoculation was repeated 2 times at intervals of 30d, and the control group was inoculated with 5 liter of KB medium (CK) by the same method. The field cultivation conditions and the water and fertilizer management are consistent, and no chemical pesticide is used any more. And (5) carrying out district yield measurement when the head stubble flowers of the honeysuckle flowers are harvested.
The results show that the use of the pseudomonas A34-9 microbial inoculum, 1 liter to 5 liters per mu, can effectively promote the length of the flower bud of the head stubble of the honeysuckle, the weight of the hundred needles and the accumulated head stubble flower yield (Table 7).
TABLE 7 growth promoting effect of Pseudomonas A34-9 on honeysuckle
Example 11 yield test of biocontrol bacteria on wheat
The preparation method of the Pseudomonas A34-9 fermentation broth is the same as in example 5.
And (3) applying the fermentation liquor of the biocontrol pseudomonas A34-9 in a wheat field with consistent growth vigor in a one-leaf one-heart period of the wheat. Inoculating biocontrol bacteria by spraying method, and applying 1L and 1.5L Pseudomonas A34-9 fermentation liquid (living bacteria concentration is 2×10) respectively 9 CFU/liter), 1 inoculation, control group, 1.5 liter KB medium (CK) using the same method. The field cultivation conditions and the water and fertilizer management are consistent, and no chemical pesticide is used any more. And (5) carrying out district yield measurement when the wheat is harvested.
The results show that the pseudomonas A34-9 can promote wheat tillering, increase wheat stem thickness and wheat yield, and has obvious promoting effect on wheat growth and yield (Table 8).
Table 8 promoting growth of Pseudomonas A34-9 on wheat
Project | A34-9 1L/mu | A34-9.5L/mu | CK |
Height of plant (cm) | 34.12 | 34.32 | 34.02 |
Tillering number | 3.36 | 3.25 | 3.05 |
Stem thickness (mm) | 3.46 | 3.19 | 2.95 |
Particle count/spike | 37.00 | 44.33 | 36.87 |
Cell yield (g/0.25 m) 2 ) | 254.30 | 257.50 | 202.33 |
Example 12 yield test of peanut by biocontrol bacteria
The preparation method of the Pseudomonas A34-9 fermentation broth is the same as in example 5.
And (3) applying the fermentation liquor of the biocontrol pseudomonas A34-9 in the peanut field with consistent growth vigor from the seedling stage to the flowering and needle-setting stage in the late ten days of 6 months. Inoculating biocontrol bacteria by spraying method, and applying 2L of Pseudomonas A34-9 fermentation liquid (living bacteria concentration is 2×10) per mu 9 CFU/liter), 1 inoculation, control group using the same method access 2 liter KB medium (CK). The field cultivation conditions and the water and fertilizer management are consistent, and no chemical pesticide is used any more.
The plant height, the number of peanut plants in a sample, the length of a first pair of branches, the number of branches within 15cm, the un-fruiting needle setting rate, the full fruit rate and the number of branches are respectively measured in the full fruit maturity period of peanuts, the fresh weight of 10 peanut pods is investigated in the peanut harvesting period, the un-fruiting needle setting rate, the occurrence condition of root rot are measured, and the pod weight and the kernel weight are measured after airing.
As can be seen from Table 9, pseudomonas A34-9 can effectively increase the number of peanut plants in unit area, reduce the plant height and the first pair of branch lengths, increase the number of sections by 15cm, and can effectively improve the peanut setting rate in theory. Meanwhile, the unoccupied needle setting rate is reduced, and the yield is measured in the final harvest period, so that the pseudomonas A34-9 can effectively improve the weight of the hundred kernels and the weight of the hundred pods and effectively improve the peanut yield (Table 9). As can be seen from FIG. 3, the peanut field growth using strain A34-9 is significantly better than that of the control group.
TABLE 9 growth promoting effect of Pseudomonas A34-9 on peanut
Project | CK | A34-9 |
Average plant height (cm) | 46.89 | 42.83 |
Average plant number in sample (plant/0.8 m) 2 ) | 24.4 | 31 |
First pair of branch lengths (cm) | 50.70 | 46.69 |
15cm section number | 3.77 | 4 |
No results were given to the needle percent | 16.15 | 12.61 |
Fruit saturation (%) | 30.26 | 30.54 |
Number of branches | 8.7 | 8.7 |
10 peanut pod fresh weight yield (g) | 283.63 | 347.50 |
Yield increase (%) | 0 | 22.52 |
Drying rate | 58.21 | 58.96 |
No results were given to the needle percent | 18.15 | 14.00 |
Fruit saturation (%) | 42.55 | 44.42 |
Weight of hundred pods | 127.35 | 130.47 |
Hundred kernel weight | 56.51 | 58.97 |
Prevention effect of root rot (%) | 0 | 52.87 |
The foregoing description is only of the preferred embodiments of the present invention, and various modifications and changes will occur to those skilled in the art. All possible modifications, equivalents, improvements, etc. which fall within the spirit and principles of the invention are intended to be included within the scope of the invention.
Claims (10)
1. Pseudomonas capable of inhibiting plant pathogenic bacteria and having growth promoting effect, characterized in that it is Pseudomonas A34-9 (Pseudomonas sp.A34-9) deposited with China center for type culture Collection, accession number: cctccc NO: m2022397, date of preservation: 2022, 04, 08.
2. The pseudomonas as claimed in claim 1 wherein said phytopathogen comprises: fusarium, rhizoctonia solani, and phytophthora.
3. The pseudomonas of claim 2, wherein the fusarium comprises: fusarium asiaticum, fusarium equisetum, fusarium graminearum, fusarium pseudograminearum, fusarium layering, fusarium putrescens, fusarium oxysporum.
4. The pseudomonas as claimed in claim 1 wherein the plant inhibiting plant pathogenic bacteria and having a growth promoting effect comprises: radix Salviae Miltiorrhizae, rhizoma Dioscoreae, rhizoma Atractylodis Macrocephalae, flos Lonicerae, semen Tritici Aestivi, and semen arachidis hypogaeae.
5. The method for screening Pseudomonas according to claim 1, wherein rhizosphere soil for tobacco planting in Alshan of inner Mongolia is collected, the soil is added into sterile water and sufficiently oscillated to obtain a soil suspension, then the soil suspension is subjected to multiple dilution, the dilution is coated on KB solid medium, the culture is inverted in a constant temperature incubator at 28 ℃ for 36 hours, and after bacterial colony grows out, the bacterial strain is separated and purified, and the bacterial strain is screened.
6. A method for preparing a fermentation broth of pseudomonas as claimed in claim 1 when applied to plant growth promoting plants, comprising the steps of: inoculating pseudomonas A34-9 into KB culture medium, and carrying out shaking culture at 28 ℃ and 180rpm for 16 hours to prepare seed bacteria; inoculating seed bacteria into KB culture medium with an inoculum size of 3% (v/v), and fermenting at 28 ℃ and 180rpm for 48 hours to form fermentation liquor; the concentration of the pseudomonas viable bacteria in the fermentation liquor is 10 9 ~10 10 CFU/mL。
7. The method of claim 6, wherein the KB medium comprises: 20g of peptone, 10mL of glycerol, 1.5g of dipotassium hydrogen phosphate, 1.5g of magnesium sulfate heptahydrate and 0g or 20g of agar are added with distilled water to a volume of 1000mL, and after uniform mixing, the mixture is packaged in triangular flasks and sterilized at 121 ℃ for 30min.
8. Use of a pseudomonas as claimed in claim 1 for inhibiting plant pathogenic bacteria.
9. Use of the pseudomonas as claimed in claim 1 for plant growth promotion.
10. Use of the pseudomonas as claimed in claim 1 for controlling soil-borne root diseases of crops.
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