Orchard soil conditioner and application thereof

Abstract

The invention belongs to the technical field of soil conditioners, and particularly relates to an orchard soil conditioner and application thereof. The invention discloses a strain TXB1-10 and application of fermentation liquor of the strain TXB1-10 in prevention and treatment of nematodes, the taxonomic name of the strain TXB1-10 is Sinomonas atrocyania, the strain TXB1-10 is preserved in the China Center for Type Culture Collection on December 28, 2020, and the preservation number of the strain TXB1-10 is CCTCC No: M 2020984. When the bacterium content of fermentation liquor of the strain TXB1-10 is 1.0 * 10 < 8 > cfu / g, the LC50 values of the strain TXB1-10 to meloidogyne incognita and meloidogyne javanica are 76.3959 mg / L and 104.5214 mg / L respectively, so that the strain TXB1-10 and the fermentation liquor of the strain TXB1-10 have the potential of being developed into a soil conditioner or a nematicide for preventing and treating the orchard nematodes.

Description

Technical Field

[0001] The invention belongs to the technical field of soil conditioners, and in particular relates to an orchard soil conditioner and application thereof. Background Art

[0002] In orchards such as bananas, citrus and grapes, root-knot nematode disease caused by nematodes of the genus Meloidogyne is a disease that cannot be ignored. Root-knot nematodes are the pathogens of plant nematode diseases of the genus Meloidogyne of the order Meloidogyne. They have a wide host range, with more than 3,000 plant species, including fruit crops, cash crops, food crops, vegetable crops and ornamental plants. According to records, 90% of crop root-knot nematode diseases are caused by four common nematodes: southern root-knot nematodes, peanut root-knot nematodes, Javan root-knot nematodes and northern root-knot nematodes. The dominant species of banana root-knot nematodes is the southern root-knot nematode, which accounts for more than 50% of the population, followed by the Javan root-knot nematode, which accounts for about 20%.

[0003] At present, the prevention and control of root-knot nematode disease is mainly based on chemical agents, but with the increasing prominence of environmental problems and pesticide residues, new and friendly control agents are urgently needed. Strain TXB1-10 is a bacterium screened from the soil in the selenium-rich area of ​​Teng County. It can significantly increase the content of exchangeable selenium in the soil and has an activation effect on soil selenium. In the prior art, the patent with application number CN201811201689.9 discloses the screening and identification of the strain, the patent with application number CN202110734139.9 discloses that the strain has a selenium-enriched effect, and the patent with application number CN202310017640.2 discloses that the strain has a control effect on tomato gray mold, which can provide support for the development of biological agents for the control of tomato gray mold. In actual experiments, the inventors also found that the strain has a killing effect on nematodes. At present, there are no related reports on strain TXB1-10 and its fermentation liquid in the control of nematodes, in order to provide support for the development of soil conditioners or nematicides for the control of orchard nematodes.

[0004] The information disclosed in this background technology section is only intended to enhance the understanding of the overall background of the invention and should not be regarded as an acknowledgment or any form of suggestion that the information constitutes the prior art already known to a person skilled in the art. Summary of the invention

[0005] The purpose of the present invention is to provide an orchard soil conditioner and application thereof.

[0006] To achieve the above object, the present invention provides the following technical solutions:

[0007] The first object of the present invention is to provide a strain TXB1-10 and its fermentation liquid for use in controlling nematodes. The strain TXB1-10 is classified as Sinomonas atrocyanea and was deposited in the China Center for Type Culture Collection on December 28, 2020 with a deposit number of CCTCC No: M 2020984. The depository address is "Wuhan University, Wuhan, China.

[0008] More specifically, the preparation method of the fermentation broth of the strain TXB1-10 is as follows: a single colony of the strain TXB1-10 is inoculated into LB liquid culture medium, and cultured at 37°C and 180 r / min for 12 h, and then the bacterial content of the culture medium is adjusted to 1.0×10 8 cfu / g.

[0009] More specifically, the LB liquid culture medium formula is: 10 g of tryptone, 5 g of yeast powder, 10 g of sodium chloride, 1 L of distilled water, and the pH is adjusted to 7.0.

[0010] More specifically, the nematodes include southern root-knot nematodes, peanut root-knot nematodes, javanic root-knot nematodes and northern root-knot nematodes.

[0011] The second object of the present invention is to provide an orchard soil conditioner, which comprises the fermentation liquid of the strain TXB1-10.

[0012] More specifically, the soil conditioner also includes fluopyram, and the mass ratio of the fermentation liquid of the strain TXB1-10 to fluopyram is 1-8:16-1.

[0013] More specifically, the orchard soil conditioner is a mixture of the following ingredients, measured by mass percentage: 10-15% of fermentation liquid of strain TXB1-10, 10-30% of fluopyram, 3-8% of sodium dodecylbenzene sulfonate and the remainder of an inert filler.

[0014] Compared with the prior art, the present invention has the following beneficial effects:

[0015] (1) The fermentation broth of the strain TXB1-10 of the present invention contains 1.0×10 8 cfu / g, its LC against southern root-knot nematode and javanic root-knot nematode 50 The values ​​are 76.3959 mg / L and 104.5214 mg / L respectively. It can be seen that the strain TXB1-10 of the present invention and its fermentation liquid have the potential to be developed into a soil conditioner or nematicide for controlling orchard nematodes.

[0016] (2) Within the mass ratio range of 1-8:16-1, the mixed use of the fermentation broth of the strain TXB1-10 of the present invention and fluopyram exhibits a synergistic effect on nematodes. Compared with the use of the fermentation broth of the strain TXB1-10 or fluopyram alone, the control effect on nematodes can be improved, which can provide support for the development of orchard soil conditioners or nematicides for controlling nematodes.

[0017] Storage Information

[0018] Sinomonas atrocyanea strain TXB1-10, its preservation number is CCTCCNo: M2020984, the preservation date is December 28, 2020, the preservation unit is the China Center for Type Culture Collection, and the preservation address is Wuhan University, Wuhan, China. DETAILED DESCRIPTION

[0019] The following is a clear and complete description of the technical solution of the patent of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by technicians in this field without creative work are within the scope of protection of the present invention.

[0020] The strain TXB1-10 has been disclosed in the prior art, and this application only studies its performance in inhibiting nematodes.

[0021] Example 1: Determination of the toxicity of the fermentation broth of strain TXB1-10 to nematodes

[0022] 1.1 Test subjects

[0023] Southern root-knot nematode (Meloidogyne incognita) and Javan root-knot nematode (Meloidogyne javanica) were collected from the banana experimental field of Guangxi Academy of Agricultural Sciences.

[0024] The nematode eggs were hatched to obtain second-instar larvae of the same age, and the root-knot nematodes were resuspended in clean water to 400 heads / mL to obtain a nematode suspension for later use.

[0025] 1.2 Fermentation broth of strain TXB1-10

[0026] A single colony of strain TXB1-10 was inoculated into LB liquid medium and cultured at 37°C and 180 rpm for 12 h. The bacterial content of the culture medium was then adjusted to 1.0 × 10 8 cfu / g (i.e., fermentation broth of strain TXB1-10). The formula of LB liquid culture medium is: 10 g tryptone, 5 g yeast powder, 10 g sodium chloride, 1 L distilled water, and the pH is adjusted to 7.0.

[0027] The fermentation broth of strain TXB1-10 (100% by mass) was diluted into 5 concentration gradients by equal ratio with 0.1% Tween-80 aqueous solution for later use.

[0028] 1.3 Test method (refer to "NT / Y 1833.1-2009 Guidelines for Indoor Bioassay Tests of Pesticides - Nematicides - Part 1 - Tests for Inhibition of Plant Pathogenic Nematodes - Insect Immersion Method")

[0029] Mix 4 mL of the fermentation liquid dilution of strain TXB1-10 and 4 mL of the nematode suspension evenly, use a pipette to transfer a certain volume of the above-mentioned mixture into the small holes of a 24-well biochemical test plate, cover it, and culture it at a constant temperature of 25°C. At the same time, set the treatment of 0.1% Tween-80 aqueous solution as a blank control, and set 3 replicates for each mass concentration. After 24 hours, take 1 mL of the mixture from each treatment to observe the death of nematodes under a dissecting microscope. The number of nematodes observed in each replicate is not less than 100, record the total number of nematodes investigated and the number of dead nematodes, and calculate the mortality rate and corrected mortality rate of each treatment. The criterion for the death of nematodes is: the nematodes are stiff and cannot bend and move when touched with a hairpin or bamboo needle.

[0030]

[0031] In the above formula: P--mortality rate, unit is %; K--number of dead insects; N--total number of insects treated.

[0032]

[0033] In the above formula: P1--corrected mortality rate, unit is %; P t --Treatment mortality rate, in %; P0--blank control mortality rate, in %.

[0034] 1.4 Data Analysis

[0035] The DPS software was used to perform regression analysis on the logarithmic values ​​of the concentration of each treatment agent and the corrected mortality probability value of each treatment, and the LC value of the fermentation liquid of strain TXB1-10 on nematodes was calculated. 50 The results are shown in Table 1.

[0036] Table 1 Results of toxicity test of the fermentation liquid of strain TXB1-10 against nematodes

[0037]

[0038] As shown in Table 1, the LC of the fermentation liquid of strain TXB1-10 against southern root-knot nematodes and javanic root-knot nematodes 50 The values ​​are 76.3959 mg / L and 104.5214 mg / L respectively.

[0039] Example 2: Determination of the combined toxicity of the fermentation broth of strain TXB1-10 and fluopyram to nematodes

[0040] In the early stage, the compatibility test found that fluopyram had a strong inhibitory effect on the growth of strain TXB1-10 at a low concentration, and the compatibility was poor; while fluopyram and strain TXB1-10 had good compatibility. Therefore, fluopyram was selected to be mixed with the fermentation liquid of strain TXB1-10.

[0041] 2.1 Test subjects

[0042] Southern root-knot nematode (Meloidogyne incognita)

[0043] 2.2 Test agents

[0044] 1.0×10 8 cfu / g strain TXB1-10 fermentation broth (same as 1.2), 96% Fluopyram technical

[0045] The fermentation liquid of strain TXB1-10 (100% by mass) is diluted into a single-dose mother liquor with a 0.1% Tween-80 aqueous solution; the fluopyram original drug is first dissolved with dimethyl sulfoxide, and then diluted with a 0.1% Tween-80 aqueous solution to form a single-dose mother liquor. Multiple groups of proportions are set up, and each single-dose mother liquor and each group of proportioned mixtures are diluted with a 0.1% Tween-80 aqueous solution to 5 mass concentration gradients for standby use. The above agents are prepared and used immediately.

[0046] 2.3 Test methods

[0047] Mix 4 mL of the drug solution and 4 mL of the nematode suspension evenly, use a pipette to transfer a certain volume of the above mixture into the small holes of a 24-well biochemical test plate, cover it, and culture it at a constant temperature of 25°C. At the same time, set the treatment of 0.1% Tween-80 aqueous solution as a blank control, and set 3 replicates for each mass concentration. After 24 hours, take 1 mL of the mixture from each treatment to observe the death of nematodes under a dissecting microscope. The number of nematodes observed in each replicate is not less than 100, record the total number of nematodes investigated and the number of dead nematodes, and calculate the mortality rate and corrected mortality rate of each treatment.

[0048] 2.4 Data Analysis

[0049] DPS software was used to perform regression analysis on the logarithmic values ​​of the concentration of each treatment and the corrected mortality probability value of each treatment to calculate the LC value of the drug solution on nematodes. 50 The synergistic effect was evaluated according to the Wadlley method.

[0050] SR ≥ 1.5 indicates a synergistic effect; SR ≤ 0.5 indicates an antagonistic effect; 0.5 < SR < 1.5 indicates an additive effect. The results are shown in Table 2.

[0051] Table 2 Combined toxicity test of the fermentation broth of strain TXB1-10 and fluopyram against southern nematodes

[0052]

[0053] As shown in Table 2, within the mass ratio range of 1-8:16-1, the mixed use of the fermentation broth of strain TXB1-10 and fluopyram showed a synergistic effect on southern root-knot nematodes. Compared with the use of the fermentation broth of strain TXB1-10 or fluopyram alone, the control effect on southern root-knot nematodes can be improved, which can provide support for the development of soil conditioners or nematicides for controlling nematodes.

Claims

1. Use of a strain TXB1-10 and its fermentation liquid in controlling nematodes, characterized in that: The strain TXB1-10 has the scientific name Sinomonas atrocyanea and was deposited in the China Center for Type Culture Collection on December 28, 2020, with the collection number CCTCC No: M 2020984.

2. The use according to claim 1, characterized in that: The preparation method of the fermentation broth of the strain TXB1-10 is as follows: a single colony of the strain TXB1-10 is inoculated into an LB liquid culture medium, and the culture is shaken and cultured at 37°C and 180 r / min for 12 hours, and then the bacterial content of the culture medium is adjusted to 1.0×10 8 cfu / g.

3. The use according to claim 1, characterized in that: The LB liquid culture medium formula is: 10 g of tryptone, 5 g of yeast powder, 10 g of sodium chloride, 1 L of distilled water, and the pH is adjusted to 7.

0.

4. The use according to claim 1, characterized in that: The nematodes include southern root-knot nematodes, peanut root-knot nematodes, Javan root-knot nematodes and northern root-knot nematodes.

5. An orchard soil conditioner, characterized in that: The orchard soil conditioner comprises the fermentation liquid of the strain TXB1-10 according to claim 1.

6. The orchard soil conditioner according to claim 5, characterized in that: The soil conditioner also includes fluopyram, and the mass ratio of the fermentation liquid of the strain TXB1-10 to fluopyram is 1-8:16-1.

7. The orchard soil conditioner according to claim 6, characterized in that: The orchard soil conditioner is prepared by mixing the following ingredients by mass percentage: 10-15% of fermentation liquid of strain TXB1-10, 10-30% of fluopyram, 3-8% of sodium dodecylbenzene sulfonate and the remainder of inert filler.

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