Bacillus toyonensis strain as well as preparation method and application of microbial agent thereof

A technology of Bacillus orientalis and inoculum, which is applied in the field of microbial isolation and culture, and achieves the effects of simple and easy preparation method, strong environmental adaptability and environmental friendliness, and low cost

Active Publication Date: 2018-03-13
YUNNAN ACAD OF TOBACCO AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, according to the available data, the preparation and utilization of Bacillus orientalis and its preparations that can simultaneously prevent and treat tobacco black shank and root-knot nematode have not been reported. The present invention provides biocontrol for preventing and treating these two diseases in tobacco. resources and methods

Method used

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preparation example Construction

[0045] Oriental bacillus described in the present invention ( Bacillus toyonensis ) The preparation method of the bacterial agent of SB3018, including the steps of slant culture, seed culture, fermentation and bacterial agent preparation, specifically including:

[0046] A, slant culture: Bacillus orientalis ( Bacillus toyonensis ) SB3018 was inoculated on the slant medium, and cultured at a temperature of 28-30°C for 48 hours to obtain slant strains;

[0047] B. Seed culture: inoculate the slant strains into the liquid medium, and cultivate them at a temperature of 28-32°C for 18-24 hours to obtain liquid-fermented seeds;

[0048] C, fermentation: the liquid fermented seeds are inoculated into the fermenter according to the amount of 3~5% of the volume of the fermentation medium, cultivated at 28~34°C for 36~48h, the stirring speed is 120~150rpm, and the ventilation rate is 5~10%, to obtain Bacillus orientalis SB3018 fermentation broth;

[0049] D. Preparation of bacteri...

Embodiment 1

[0056] The identification method of SB3018 provided by the invention.

[0057] The PCR amplification and sequencing of the 16SrDNA gene of SB3018 were completed by Shanghai Yingjun Biotechnology Co., Ltd. A phylogenetic tree was constructed based on the sequencing results. Phylogenetics showed that strains with Bacillus The evolutionary distance of sp. is the closest. Combining the morphology, colony characteristics and physiological and biochemical characteristics, the strain is identified as Bacillus orientalis ( Bacillus toyonensis ).

Embodiment 2

[0059] Toxic effect of liquid preparation of SB3018 bacteria on root-knot nematodes

[0060] The isolation of root-knot nematodes was carried out by the Behmann funnel method, and the second instar larvae of root-knot nematodes were obtained. The isolated root-knot nematodes were added to a 100-fold dilution of the liquid preparation of Bacillus tonicii SB3018, and the same amount of water was used as a control, and the nematodes were left at room temperature for 1 hour to observe the number of surviving and dead nematodes. The results show that, in the control, the mortality rate of the second instar larvae of root-knot nematode is 4.41%, and the mortality rate of the second instar larvae of root-knot nematode in the treatment of the 100-fold dilution of the liquid preparation of Bacillus orientalis SB3018 reaches 100%, thus It can be seen that the liquid preparation with Bacillus orientalis SB3018 as the active ingredient has a strong poisonous effect on the second instar la...

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Abstract

The invention discloses a bacillus toyonensis strain as well as a preparation method and application of a microbial agent thereof. The preservation department of the bacillus toyonensis SB3018 is China General Microbiological Culture Collection Center (CGMCC), the preservation date of the bacillus toyonensis SB3018 is November 21, 2014, the preservation number of the bacillus toyonensis SB3018 isCGMCC No. 10033; the bacillus toyonensis belongs to the realm of eubacteria, firmicutes, bacilli, bacillaceae and bacillus. The microbial agent of the bacillus toyonensis SB3018 is prepared by carrying out slant culture, seed culture and fermentation on the bacillus toyonensis SB3018. The preparation method comprises the steps of slant culture, seed culture, fermentation and microbial agent preparation; the microbial agent of the bacillus toyonensis SB3018 is applied to preparation of medicines for preventing and controlling tobacco root knot nematode disease and tobacco black shank. The preparation prepared by taking the bacillus toyonensis as an active component has a remarkable killing effect for the tobacco root knot nematode.

Description

technical field [0001] The invention belongs to the technical field of microorganism isolation and cultivation, and in particular relates to a strain of Bacillus orientalis and a preparation method and application of a bacterial agent. Background technique [0002] Tobacco is an important economic crop, and its growth process is harmed by many diseases. Tobacco root-knot nematode and black shank are two important soil-borne diseases, which cause tobacco yield reduction or failure. root-knot nematodes ( Meloidogyne spp.) is a class of plant endoparasitic nematodes, without strict host specificity, can infect a variety of crops, seriously endanger agricultural production, and cause global losses of about 100 billion US dollars every year. It is widely distributed all over the world and has a wide range of hosts. It can make it easier for bacteria and fungi to infect plants, and is one of the important reasons for inducing plant diseases. With the development of tobacco prod...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/00A01P5/00A01P3/00C12R1/07
CPCA01N63/00C12N1/20C12N1/205C12R2001/07
Inventor 夏振远马雁军卢灿华莫笑晗王学坚余清许银莲
Owner YUNNAN ACAD OF TOBACCO AGRI SCI
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