Bacillus thuringiensis and application thereof

A technology of Bacillus aureus and strains, applied in the field of biological control, can solve the problems of difficult practical application and poor control effect in the field, and achieve the effects of good drug effect persistence, strong specialization and high control effect

Active Publication Date: 2018-01-19
INST OF PLANT PROTECTION HEBEI ACAD OF AGRI & FORESTRY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] After retrieval, in terms of biological control, there are only reports on the use of entomopathogenic nematodes to control Flea beetle oligochaete Very sensitive and greatly affected by the environment, the field control effect is not good, and it is difficult to apply in practice
So far, no reports have been found on the use of Bacillus thuringiensis to control flea beetles

Method used

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  • Bacillus thuringiensis and application thereof
  • Bacillus thuringiensis and application thereof
  • Bacillus thuringiensis and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Embodiment 1 The process of collection and screening of bacterial strain DK49 of the present invention

[0040]In August 2015, the Institute of Plant Protection, Hebei Academy of Agriculture and Forestry Sciences collected more than 2,000 soil samples in Hebei Province, and isolated more than 100 Bt strains from them. More than 100 isolated Bt strains were mixed in 30mL LB liquid medium (composed of: tryptone 10g, yeast extract 5g, sodium chloride 10g, add water to 1L), at 30°C, 220rpm Cultivate under conditions until the cell crystals are separated, and after counting, transfer the bacterial suspension to a 50mL centrifuge tube, centrifuge at 4°C and 8000rpm for 5min, and collect the bacterial cells; resuspend the bacterial cells with sterile water to reach a final concentration of 1×10 7 spores / mL. The indoor biological activity of Flea beetle adults was measured by dip feeding method, and finally a Bt strain with high insecticidal activity against Flea beetle was ob...

Embodiment 2

[0041] Example 2 According to the morphological characteristics, the bacterial strain DK49 of the present invention is initially classified and identified

[0042] Inoculate the DK49 strain on LB solid medium (its composition and ratio are: tryptone 10g, yeast extract 5g, sodium chloride 10g, agar powder 15g, add water to 1L), and culture at 30°C , Sampling at different times for microscopic examination to observe the morphological characteristics and crystal characteristics of the colonies. Observation results at different culture stages on LB medium are as follows: vegetative bodies: rod-shaped, blunt at both ends, about 1.1-1.5×0.5 μm in size; single or more than two exist in chains. Spores: oval, about 1.0-1.2×0.5μm in size, dormant; have strong resistance to adverse environments such as high temperature or dryness. Paraspora crystals: spherical, about 0.4 μm in diameter, with insecticidal activity. These morphological characteristics are basically consistent with the mo...

Embodiment 3

[0043] Example 3 Classification and Identification of DK49 at the Molecular Level

[0044] (1) Identification and classification of DK49 strains using gyrB gene sequence:

[0045] Inoculate the DK49 strain in LB culture medium, shake overnight at 30°C, take 1mL of the bacterial liquid into an Eppenddorf tube, centrifuge, discard the supernatant, suspend with 200μL ultrapure water, treat at 100°C for 10min, centrifuge at 12000rpm for 5min, and take Qing is the DNA template of the DK49 strain. The general primers gyrB-F and gyrB-R were used as primers for PCR amplification to obtain PCR amplification products; wherein the primers were: gyrB-F: 5'-TTGRCGGHRGYGGHTATAAAGT-3', gyrB-R: 5'-TCCDCCSTCAGARTCWCCCTC -3'. Wherein the PCR reaction system (50μL) is: 10×PCR Buffer (Mg 2+ ) 5 μL; dNTPMixture (2.5mM) 5 μL, primers (10 μmol / L) 1 μL each, template 1 μL, Taq polymerase (5U / μL) 1 μL, and ultrapure water to 50 μL. The PCR reaction program was: 95°C for 5min; 30 cycles of 95°C for...

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Abstract

The invention discloses a bacillus thuringiensis strain DK49 which belongs to the field of biological control and has the preservation number of CGMCC No.14761. The invention also discloses a microbial insecticide containing DK49 and application thereof in control of Luperomorpha insect pests. The bacillus thuringiensis DK49 disclosed by the invention has a control effect on Luperomorpha suturalisChen reaching 96.81%, specificity on the Luperomorpha suturalis Chen is strong, and biocidal effect duration is good; secondly, the microbial insecticide belongs to a natural product and is non-toxicto people and livestock and pollution-free to the environment; and the microbial insecticide also has the advantages of simple preparation method and convenience in use.

Description

technical field [0001] The invention belongs to the field of biological control, and in particular relates to a bacillus thuringiensis, and also relates to the use of the bacterium in the control of vegetable pests. Background technique [0002] Luperomorpha suturalis belongs to the genus Luperomorpha suturalis of Coleoptera Chrysomorphaceae, commonly known as "jumping insect", which mainly harms economic crops such as green onions, onions, leeks and garlic, and has become one of the main pests of leeks. Adults and larvae of Flea beetle oligochaete can be harmful. The larvae feed on the internal tissues of the fibrous roots of leeks, resulting in a decrease in the ability of the roots to absorb water and inorganic salts. even death. Adults feed on the leaves above the ground, causing nicks or holes, and black feces are attached to them, seriously affecting the yield and quality of leeks. [0003] At present, the control methods for Flea beetle oligochaete are mainly physic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12Q1/689C12Q1/04C12N15/11A01N63/02A01P7/04C12R1/07
Inventor 宋健杜立新曹伟平
Owner INST OF PLANT PROTECTION HEBEI ACAD OF AGRI & FORESTRY SCI
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