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Trans-aconitic acid producing bacteria and application thereof

A technology for trans-aconitic acid and bacteria production, which can be used in applications, bacteria, biocides, etc., and can solve problems such as hidden hazards and difficult prevention and control.

Active Publication Date: 2013-12-04
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Phytopathogenic nematodes mainly live in the soil or parasitize in plants, causing hidden harm and difficult control

Method used

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  • Trans-aconitic acid producing bacteria and application thereof
  • Trans-aconitic acid producing bacteria and application thereof
  • Trans-aconitic acid producing bacteria and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Example 1: Isolation, screening and identification of trans-aconitic acid producing bacteria YBT-1501

[0016] In this example, two trans-aconitic acid-producing bacteria were obtained from Bacillus thuringiensis YBT-1501 and Bacillus thuringiensis YBT-1520. The specific screening steps are as follows:

[0017] 1. Isolation and screening of Bacillus thuringiensis YBT-1501

[0018] Acetate selective medium (abbreviated as BPA medium) is used to separate from the soil of Huazhong Agricultural University in Shizishan District, Hongshan District, Wuhan City, Hubei Province, China, and use high performance liquid chromatography (HPLC) to detect TAA, and obtain the present invention through screening. strain.

[0019] Screening and isolation media are as follows:

[0020] 1) BPA medium formula: beef extract 5g, peptone 10g, sodium acetate 34g; add distilled water to 1L, adjust the pH value to 7.2-7.4.

[0021] 2) Separation method

[0022] Weigh 1g of the soil sample ment...

Embodiment 2

[0035] One is TAA standard product (purchased from TCI company, company website: www.tcichemicals.com / zh / cn), with a purity of 98%, which is recorded as TAA-1. The other is the crude pure product of TAA extracted from the fermentation broth of Bacillus thuringiensis YBT-1501 according to the method in Example 1, which is designated as TAA-2. Example 2: Bioassay of trans-aconitic acid (TAA) on root-knot nematode (Meloidogyne incognita)

[0036] The roots of tomato plants infested by Meloidogyne incognita were collected from the greenhouse of Huazhong Agricultural University in Hongshan District, Wuhan City, Hubei Province, China. The tomato roots were washed with sterile distilled water, and the egg masses of the above-mentioned root-knot nematode were removed from the roots and placed on ice. In the petri dish above, use 1% H in the aseptic operation bench 2 o 2 And 0.025% KI disinfection 2 times, each 15min. Transfer the sterilized egg mass to another sterilized petri dish...

Embodiment 3

[0042] Example 3: Bioassay of trans-aconitic acid (TAA) on soybean cyst nematode (Heterodera glycine Ichinohe)

[0043] Isolation of cysts by floating method from soil where soybean cyst nematode (Heterodera glycine Ichinohe) is seriously infested, and the cysts were placed in ZnSO 4 Soak in the solution for 24 hours, and after the soybean cyst nematode larvae hatch at room temperature, dilute until there are about 200 2nd instar larvae in every 1ml suspension under microscope inspection, and set aside.

[0044] The hatched 2nd instar soybean cyst nematode (Heterodera glycine Ichinohe) was used as the target organism of the bioassay. Bioassay of virulence was carried out on a commonly used 24-well plate: about 200 2nd-instar soybean cyst nematode (Heterodera glycine Ichinohe) larvae were inhaled into each well, TAA with known concentration was added (see Table 2 for specific concentration), and blank treatment was added The same amount of sterile deionized water, with a total...

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Abstract

The invention belongs to the field of microbe biotechnology and relates to the field of biological pesticides. Two trans-aconitic acid producing bacteria, that is, bacillus thuringiensis YBT-1501 and bacillus thuringiensis YBT-1520 are obtained from bacillus thuringiensis through screening; through utilizing the fermentation products of the two strains, the coarse and refine product of the trans-aconitic acid can be obtained; according to bioassay, the trans-aconitic acid has the activity of poisoning and killing meloidogyne incognita, heterodera glycine ichinohe and ditylenchus destructor, and the activity of suppressing the growth of cotton bollworm and housefly. The trans-aconitic acid producing bacteria are novel biological disinsection pesticides and a novel prevention and cure way for the current prevention and cure strategy is provided. The bacillus thuringiensis YBT-1501 which is used for preparing the trans-aconitic acid is preserved in the China Center for Type Culture Collection, with the serial number of CCTCC NO: M2012236.

Description

technical field [0001] The invention belongs to the field of biological pesticides. It specifically relates to a trans-aconitic acid (TAA)-producing bacterium, the fermentation liquid of which is toxic and active against root-knot nematode incognita, soybean cyst nematode, and sweet potato stem nematode, and is effective against cotton bollworm and housefly It has the activity of inhibiting growth, and can be used to prevent and control agricultural pests such as plant pathogenic nematodes which are extremely harmful to agricultural production. Background technique [0002] Nematodes belong to the phylum Nematode of the kingdom Animalia and are a class of bilaterally symmetrical protocoelomate invertebrates. There are many kinds of nematodes, which are second only to insects in terms of species and quantity. About 10% of nematodes parasitize on plants, which are called plant-parasitic nematodes (PPN). Phytopathogenic nematodes are small, transparent insects that are usuall...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P7/48A01N63/02A01P5/00A01P7/04C12R1/07
Inventor 孙明曹诗云都萃颖聂相涛施祥雨彭东海阮丽芳
Owner HUAZHONG AGRI UNIV
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