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Bacillus megatherium and application thereof

A technology of Bacillus megaterium and strains, applied in the field of Bacillus megaterium and its application in the control of rice nematodes, can solve the problems of high toxicity and long residual time, and achieve excellent nematicidal activity

Active Publication Date: 2019-06-11
HUNAN AGRICULTURAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In the prior art, non-fumigating nematicides such as organophosphorus or carbamates are mostly used, such as carbofuran, fenamiphos and other nematicides with high toxicity, long residual time, and easy to pollute the environment to prevent and control rice nematodes

Method used

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  • Bacillus megatherium and application thereof
  • Bacillus megatherium and application thereof
  • Bacillus megatherium and application thereof

Examples

Experimental program
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preparation example Construction

[0024] The present invention also provides a preparation method of the preparation described in the above scheme, the steps are: inoculating Bacillus megaterium D45 into a fermentation medium for fermentation to obtain the preparation. In the present invention, there is no special limitation on the inoculation amount of the Bacillus megaterium D45; the temperature of the fermentation is preferably 25-30°C, more preferably 26-29°C, most preferably 27-28°C; the fermentation time It is preferably 36-72 hours, more preferably 48 hours; the fermentation method is preferably shaker fermentation; the rotation speed of the shaker is preferably 150-200 r / min, more preferably 180 r / min.

[0025] In the present invention, after fermentation, it is preferred to also centrifuge the fermented liquid obtained from the fermentation; the rotational speed of the centrifugation is preferably 5000-10000r / min, more preferably 8000r / min; the time of the centrifugation is preferably 8 to 15 minutes,...

Embodiment 1

[0028] Example 1 Isolation and cultivation of Bacillus megaterium strain (Bacillus megaterium) D45 (CGMCC15119)

[0029] Soil sample dilutions were prepared by the equal dilution method. Weigh 2g of soil and pour it into a beaker filled with 30mL sterile water. After shaking, the gradient (10 -1 -10 -7 ) was diluted, and 100 μL of the dilutions in the last three dilution gradient test tubes were applied to the beef extract peptone culture plate, dried on the ultra-clean bench, and the coated plate was incubated at 28°C for 2 days in the dark.

[0030] Pick the isolated bacterial single colony with a sterilized toothpick onto the medium for purification to obtain isolated and purified bacteria. The isolated and purified bacteria were transferred into a round-bottomed glass test tube (20×200 mm) with a sterilized toothpick, and cultured at 28° C. and 180 r / min for 48 hours to obtain a fermentation broth.

Embodiment 2

[0031] Embodiment 2: the strain of purification obtained in embodiment 1 is identified

[0032] (1) Morphological characteristics

[0033] Colonies cultured on beef extract peptone medium at 25°C±1°C for 24h have irregular morphology, wrinkled edges, rough and opaque, dry surface, milky white or beige, see figure 1 . Observed under an optical microscope, the bacterial body of the D45 strain was rod-shaped with a round end and flagella all over the body.

[0034] (2) Physiological and biochemical experiments

[0035] The physiological and biochemical characteristics of biocontrol bacteria D45 are Gram reaction positive, can hydrolyze starch, can produce reduction reaction with nitrate, methyl red test is negative reaction, can use catalase, does not produce hydrogen sulfide, has The ability to decompose gelatin, the acetylmethylmethanol test was positive.

[0036] (3) 16SrDNA sequencing and sequence analysis

[0037] Bacterial genomic DNA was extracted using bacterial univ...

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Abstract

The invention provides a bacillus megatherium strain D45. The bacillus megatherium strain D45 has a preservation number of CGMCC No.15119, and belongs to the technical field of plant nematode biological prevention and control. A fermentation product of the bacillus megatherium strain D45 has excellent nematode killing activity and can be used for preventing and controlling Meloidogyne graminicolaand Heterodera elachista. Proved by verification, after a fermentation broth of the bacillus megatherium strain D45 is diluted by 2 folds and 5 folds, the root knot inhibition rates on Meloidogyne graminicola are 67.3% and 62.6% respectively, and the prevention and control effects are 53.1% and 56.3% respectively. The prevention and treatment effect of the D45 strain fermentation broth 2-fold diluent on Heterodera elachista is 60%.

Description

technical field [0001] The invention relates to the technical field of biological control of plant nematodes, in particular to a bacillus megaterium and its application in the control of rice nematodes. Background technique [0002] Plant nematodes that parasitize rice roots are an important class of pathogenic nematodes that cause rice root diseases, and usually cause rice yields to decrease by more than 10%. The nematodes that harm my country's rice mainly contain root-knot nematode (Meloidogynegraminicola), cyst nematode (Heterodera elachista), rice stem-point nematode (Aphlenchoides besseyi) and root-knot nematode (Hirschmanniellaspp.) etc., wherein root-knot nematode (Meloidogyne graminicola) M. graminicola) is considered to be a serious threat to rice plant parasitic nematodes. [0003] Root-knot nematodes of the grass family are mostly distributed in tropical and subtropical regions, and occur in Guangdong, Guangxi, Hainan, Yunnan, Fujian and other places in my countr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/02A01P5/00C12R1/11
Inventor 丁中叶姗
Owner HUNAN AGRICULTURAL UNIV
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