Bacillus thuringiensis bacterial strain for strain insect disinfestations, restraining epiphyte and uses thereof

A technology of Bacillus chrysogenum and strains, applied in the field of biological control of plant diseases and insect pests, can solve the problem of not being able to kill beet armyworm with high efficiency, and achieve the effect of inhibiting the germination of fungal spores

Inactive Publication Date: 2008-08-20
NANKAI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

None of the above-mentioned strains have the characteristics of highly effective killing beet armyworm

Method used

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  • Bacillus thuringiensis bacterial strain for strain insect disinfestations, restraining epiphyte and uses thereof
  • Bacillus thuringiensis bacterial strain for strain insect disinfestations, restraining epiphyte and uses thereof
  • Bacillus thuringiensis bacterial strain for strain insect disinfestations, restraining epiphyte and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Embodiment 1, the screening of multifunctional bacterial strain Bt 519-1 and the determination of antifungal activity

[0022] 1.1. Collect soil samples, use the heat-resistant principle of Bacillus, treat the samples at 80°C for 20 minutes, observe with a microscope after cultivation, and select a strain with spores and crystals, named Bt 519-1.

[0023] 1.2. A pair of specific primers were designed according to the chiB gene sequence released by GenBank, and the PCR detection of the 70kDa chitinase gene was carried out. The Bt 519-1 strain was positive for the chiB gene of chitinase.

[0024] 1.3. Prepare a solid medium with colloidal chitin as the only carbon source, and its formula is (g / L): colloidal chitin 20, KH 2 PO 4 1, Na 3 PO 4 2. MgSO 4 ·7H 2 O 0.5, KCl 0.5, agar powder 15, pH 7.2. Used to detect chitinase activity.

[0025] 1.4. Spot the Bt 519-1 strain on the above-mentioned medium with a sterile toothpick, and incubate at 30°C for 5-7 days. The ...

Embodiment 2

[0031] Example 2, Bt519-1 Determination of Inhibitory Activity to Fungal Spore Germination

[0032] The inhibitory effect of strain Bt 519-1 on fungal spore germination was detected by Oxford cup-Plate method. The specific method is:

[0033] 2.1. Take the prepared PDA plates, and place 2 sterile Oxford cups on each plate. One of them is a test cup and the other is a control cup. The Oxford cup is made of stainless steel, with a height of 10mm, an outer diameter of 8mm, and an inner diameter of 6mm.

[0034] 2.2. Get the spore suspension 20ul (contain about 10 3 spores), added to the entire Oxford glass.

[0035] 2.3. Add 180uL Bt 519-1 cell suspension (OD 600 =1.530), and 180uL sterile water was added to the control cup.

[0036] 2.4. Place the plate in an incubator at 28°C and culture it for 2-3 days to observe the effect of inhibiting spore germination. The test cup with Bt 519-1 bacterial liquid and fungal spores had no fungal growth phenomenon, which proved that th...

Embodiment 3

[0037]Embodiment 3, Bt519-1 bacterial strain to cotton bollworm and beet armyworm half-lethal concentration (LC 50 ) determination

[0038] 3.1. In order to accurately calculate the semi-lethal concentration of the screened target strain Bt 519-1 on cotton bollworm and beet armyworm, the strain was cultured on ordinary NB solid medium, and after culturing at 30°C for 72 hours, the collected spores contained And the lawn of crystal protein is made into dry powder by freeze-drying method.

[0039] 3.2. The patent strain Bt15A3 (ZL 01104421.7) preserved in our laboratory and highly effective against cotton bollworm and beet armyworm was treated as a reference strain in the same way.

[0040] 3.3. Weigh the above-mentioned freeze-dried powder with the same weight, serially dilute it into 5 concentrations respectively, and prepare infected feed with corresponding concentrations.

[0041] 3.4. Inject newly hatched larvae of cotton bollworm and beet armyworm respectively, feed 48 l...

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Abstract

The invention relates to an insecticidal and antifungal bacillus thuringiensis bacterial strain and the application. The bacterial strain is bacillus thuringiensis 519-1, which is called as Bt 519-1 for short, the collection number is CGMCC No. 2327, the bacterial strain carries the insecticidal protein coding genes of cry1Aa, cry1Ab, cry1Ac, cry1I, cry2 and the vip3A and 70 kDa chitinase coding gene chiB. The semi-lethal concentration (LC50) to spodoptera exigua of the spore crystal complex freeze-dried powder of the bacterial strain is only 8.8Mug/mL, and the bacterial strain can inhibit the growth of 8 common plant pathogen fungal myceliums and inhibit the germination of the fungal spores completely. The usage of the bacterial strain can produce the insecticidal and anti-disease multifunctional biological pesticides, which is used for the prevention and the treatment of lepidoptera major pest spodoptera exigua and bollworm; at the same time, the bacterial strain also has the inhibition function of the plant pathogen fungi.

Description

【Technical field】: [0001] The invention belongs to the technical field of biological control of plant diseases and insect pests, and in particular relates to a screening method for bacillus thuringiensis which not only kills lepidopteran pests but also inhibits plant pathogenic fungi, and a preparation method for active bacterial strains. 【Background technique】: [0002] Beet armyworm belongs to Lepidoptera Noctuidae. It is a kind of omnivorous pest with strong fecundity. It can feed on hundreds of species of host plants. It is an important pest that harms wheat, cotton and various vegetables. Bring huge loss to my country's agricultural production every year. Beet armyworm is not sensitive to biopesticides. Many scholars at home and abroad use the method of compounding biopesticides and chemical pesticides. Although it has a certain control effect, the harm caused by chemical pesticides to humans and the resistance of pests to chemical pesticides are constantly increasing. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01P3/00A01P7/04C12R1/07
Inventor 陈月华韩苗苗蔡峻谢池楚肖亮刘东
Owner NANKAI UNIV
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