Fermentation method and applications of paecilomyce lilacinus microsclerotia

A technology of Paecilomyces lilacinus and fermentation method, applied in the field of biological preparations, can solve the problems of short shelf life, poor resistance to stress, inconvenient storage and transportation, etc., and achieve the effects of long shelf life, low production cost and short fermentation period

Active Publication Date: 2015-02-18
CHONGQING UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to address the above problems, to provide a method for liquid fermentation and cultivation of Paecilomyces lilacinus microsclerotia that can be produced on a large scale and its application, and to develop Paecilomyces lilacinus microsclerotias with low production cost, high infection activity, and strong stress resistance. The new infestation structure of Paecilomyces lilacinus provides a biocontrol agent for the prevention and treatment of nematode diseases, provides high-quality mother drugs for the creation of insecticidal fungal preparations, and solves the problem of poor stress resistance of traditional spore agents of Paecilomyces lilacinus, Common technical problems of short shelf life and inconvenient storage and transportation

Method used

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  • Fermentation method and applications of paecilomyce lilacinus microsclerotia

Examples

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Effect test

Embodiment 1

[0022] Example 1 Paecilomyces lilacinus Microsclerotia Induction Culture Solution Formula Optimization and Induction Process

[0023] 1. Preparation of Paecilomyces lilacinus inoculum suspension:

[0024] Inoculate the conidia of Paecilomyces lilacinus strain (from Chongqing Engineering Research Center of Insecticidal Fungus Pesticide) onto PDA plates, cultivate them for 12 days at 25°C, and sterilize with 0.1-0.5% Tween-80 Wash the mature conidia on the plate, adjust the concentration of conidia, and prepare 1.0×10 8 spores / ml of spore suspension.

[0025] 2. Formula screening of induction culture medium

[0026] 1) Inorganic salt ion concentration:

[0027] to add KH 2 PO 4 :4.0g / L, CaCl 2 2H 2 O:0.8g / L, MgSO 4 ·7H 2 O:0.6g / L, CoCl 2 ·6H 2 O: 37mg / L, MnSO 4 ·H2O:16mg / L, ZnSO 4 ·7H 2 O:14mg / L is used as the inorganic salt ion of cultivating micro-sclerotia.

[0028] 2) Selection of carbon and nitrogen sources

[0029] Glucose with a concentration of 10g / L was ...

Embodiment 2

[0037] Embodiment 2 Morphological and structural observation of Paecilomyces lilacinus microsclerotia

[0038] Paecilomyces lilacinus strain was treated with SDA Ⅱ Liquid medium, 25~28°C, 200~250rpm for shaker culture, microscopic observation of the microsclerotia production status in the liquid medium: 1-2 days after inoculation, the conidia germinated to form hyphae, and the bacterial liquid The color did not change, but the viscosity of the culture medium and the biomass of the bacteria gradually increased; after 3 days of cultivation, the color of the bacteria began to turn brown. At this time, the amount of bacteria in the bacteria suspension continued to increase, and the front end of some mycelium began to expand, which was obviously thickened ; Observed under an optical microscope, it was found that the hyphae began to aggregate. After cultivating for 4 days, the brown color of the fermented bacterium liquid continued to deepen, and a sample was taken and observed und...

Embodiment 3

[0039] Example three The sporulation ability analysis of Paecilomyces lilacinus microsclerotia

[0040] Paecilomyces lilacinus strains were treated with SDA Ⅰ 、SDA Ⅱ 、SDA Ⅲ 、SDA Ⅳ The liquid culture medium is cultured on a shaker at 23-28° C. and 200-250 rpm. After 6 days, 5% diatomaceous earth is added to the fermented culture liquid, dried at 35° C. for 24-48 hours, and crushed. Afterwards, the mixture was respectively passed through a 60-mesh standard sieve to separate the micro-sclerotia from the diatomite, and the micro-sclerotia were sealed and stored at room temperature.

[0041] Get 10 mg of the micro-sclerotia obtained from the culture of the four culture media and evenly inoculate them on the water agar plate. After culturing for 14 days at 25°C, wash the mature conidia on the plate with 0.5% Tween-80 sterilized water. Hemocytometer count, found SDA Ⅰ Combination cultured microsclerotia can produce 3.7×10 per mg 5 spores, SDA Ⅱ Combination cultured microsclero...

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Abstract

The invention discloses a fermentation method and applications of paecilomyce lilacinus microsclerotia. The fermentation method comprises the following steps: preparing paecilomyce lilacinus spore suspension; adding the paecilomyce lilacinu spore suspension into induction culture solution, and performing shake cultivation; and filtering the fermentation liquor to obtain microsclerotia precipitate. The generated microsclerotia can adapt to outside poor environments such as high temperature, drought, strong ultraviolet, and is stress-tolerant and storage-resistant, long in shelf life and capable of continuously controlling harm; the microsclerotia is short in fermentation period, low in production cost, and strong in technological controllability, and the raw materials can be easily available. The microsclerotia can be used as an active ingredient for preparing a novel insecticide fungus preparation.

Description

technical field [0001] The invention belongs to the field of biological preparations, and in particular relates to a fermentation method and application of Paecilomyces lilacinus microsclerotia. Background technique [0002] Plant parasitic nematodes such as root-knot nematode incognita, root-knot nematode northern and peanut root-knot nematode have a wide range of parasites and often damage more than 30 kinds of vegetables such as melons, solanaceous fruits, beans, radishes, carrots, lettuce, and cabbage. It can spread some fungal and bacterial diseases, which is a major threat to the safe production of vegetables in my country, resulting in a large reduction in yield and quality of vegetable crops. At present, the prevention and control of nematode diseases mainly relies on chemical control. Extensive application of chemical pesticides leads to soil environmental pollution caused by nematode diseases, pesticide residues in agricultural products seriously exceed the standard...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14A01N63/04A01P5/00C12R1/79
CPCA01N63/30C12N1/14
Inventor 王中康宋章永殷幼平
Owner CHONGQING UNIV
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