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Bacillus subtilis and application of bacillus subtilis in preventing and curing peony root-knot nematode

A technology for Bacillus subtilis and peony root-knot nematodes, applied in the directions of application, nematicides, bacteria, etc., can solve the problems of difficult colonization, poor control effect, etc., and achieves novel strains, remarkable effects, and high content of live bacteria. Effect

Active Publication Date: 2014-11-05
SHANDONG FOREST SCI RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the existing biological agent for preventing and treating tomato root-knot nematode is not easy to colonize on peony, and the control effect is not good.

Method used

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  • Bacillus subtilis and application of bacillus subtilis in preventing and curing peony root-knot nematode
  • Bacillus subtilis and application of bacillus subtilis in preventing and curing peony root-knot nematode
  • Bacillus subtilis and application of bacillus subtilis in preventing and curing peony root-knot nematode

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Experimental program
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Effect test

Embodiment 1

[0023] Embodiment 1 Mutagenesis screening and identification of Bacillus subtilis strain SDBRM01

[0024] 1. Mutagenesis and screening of strains

[0025] (1) Starting strain: Bacillus subtilis SGBs-02 strain, which was isolated from greenhouse soil in Wangwu Village, Tianliu Town, Shouguang City, Shandong Province.

[0026] (2) Medium:

[0027] NA medium: beef extract 3g, peptone 10g, NaCl 5g, agar 14g, water 1000mL, pH7.3;

[0028] NB medium: beef extract 3g, peptone 10g, NaCl 5g, water 1000mL, pH7.3;

[0029] (3) Preparation of mutagen

[0030] Activate and culture the SGBs-02 strain at 28°C for 24 hours, pick 2-3 rings, inoculate them in a conical flask (250 mL) filled with 100 mL of NB culture solution, and culture with shaking at 200 r / min for 12 hours, as the seed solution, draw 2 mL of seeds solution, inoculated into NYBD culture solution, cultured with shaking for 36 hours, stored at 4°C, and used for later use.

[0031] (4) UV mutagenesis treatment

[0032] Dil...

Embodiment 2

[0046] Example 2 Verification of indoor effect of Bacillus subtilis SDBRM01 on root-knot nematode of peony

[0047] 1. Experimental method

[0048] 1.1 Preparation of root-knot nematode second-instar larvae suspension

[0049] Fresh egg masses of root-knot nematodes were picked from the roots of peony, incubated in a Beman funnel, and collected once after 24 hours for a total of 7 days. The nematode solution was diluted to contain 100 second-instar larvae / mL, and stored at 4°C for later use.

[0050] 1.2 Preparation of egg suspension

[0051] Cut peony roots with root-knot nematodes into pieces and put them into a triangular flask, add 100mL of 0.5% NaClO solution, shake vigorously for 1-2 minutes, quickly pour them into a 200-500-mesh sieve group and rinse them with tap water for 10 minutes, and collect them through a 500-mesh sieve Eggs in the net were diluted to an egg suspension with a concentration of 3000 grains / mL.

[0052] 1.3 Indoor and in vitro measurement

[00...

Embodiment 3

[0063] Embodiment 3 Preparation of Bacillus subtilis strain SDBRM01 wettable powder

[0064] (1) Transfer the preserved Bacillus subtilis SDBRM01 to the slant of the NA test tube, and place it in an incubator at 26°C for constant temperature cultivation for 2-3 days;

[0065] (2) Under sterile conditions, dilute the cultured Bacillus subtilis SDBRM01 with sterile water (add 5 mL of sterile water to the slope of each test tube), draw 1 mL, put it in NB liquid medium, and culture it on a shaking table at 28 ° C 1~2d to obtain the seed solution;

[0066] (3) The seed solution was inoculated into a large amount of fermentation medium at a ratio of 3.0% (volume ratio), the culture temperature was 28° C., the culture time was 36 h, and the ventilation ratio was 1:0.8. After the fermentation is completed, the number of bacteria can reach 8 billion / g;

[0067] (4) One or more of the Bacillus subtilis fermentation liquid matrix selected from peat, micronized calcium carbonate and ben...

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Abstract

The invention discloses bacillus subtilis and an application of bacillus subtilis in preventing and curing peony root-knot nematode. The strain is a bacillus subtilis strain SDBRM01 with the collection number of CGMCC No.8546. Experiments show that the bacillus subtilis SDBRM01 has characteristics of novel strain, high efficiency and high in vivo bacterium content. Under the in vitro condition, a fermentation broth is diluted 5 times and the diluted fermentation broth works for 48h and inhibition ratio of inhibiting nematode is 90.3%. Under the potting condition, 200 times of dosage of a bacillus subtilis SDBRM01 wettable powder has a remarkable effect of preventing and curing peony root-knot nematode, and the control efficiency reaches 72.05%. In addition, peony grows well; no phytotoxicity is generated; and crown weight is increased by 11.3g / plant. Thus, bacillus subtilis has an important application prospect in biological control of peony root-knot nematode.

Description

technical field [0001] The invention belongs to the field of microorganism application. Specifically, the present invention relates to a strain of Bacillus subtilis and its use. Background technique [0002] Peony is a famous traditional flower in my country, an important tourism brand and a city card for foreign exchanges in Heze City, Shandong Province. It is also one of the six leading industries in agriculture and has a special status and role in the economic and social development of Heze City. At present, the total area of ​​peonies in Heze City has grown to more than 100,000 mu, with an annual output value of nearly 3 billion yuan. However, due to the harm of root-knot nematode, which seriously affects the production and sales of peony, the quality and output of paeonol cortex have declined seriously. At the same time, root-knot nematode is an important quarantine object at home and abroad, which directly affects the export trade. In recent years, root-knot nematode...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/00A01N25/14A01P5/00A01G7/06C12R1/125
Inventor 王清海季延平朱文成刘幸红段春华刘慇
Owner SHANDONG FOREST SCI RES INST
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