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Solid culture medium for nematode and preparation method and application thereof

A technology of solid culture medium and culture medium, applied in the direction of animal husbandry, etc., to achieve the effect of improving efficiency, reducing cost and increasing yield

Inactive Publication Date: 2019-01-11
NANKAI UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are few reports on the addition of insect powder

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  • Solid culture medium for nematode and preparation method and application thereof
  • Solid culture medium for nematode and preparation method and application thereof
  • Solid culture medium for nematode and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1: Preparation of medium containing yellow mealworm larvae powder

[0034] 1.1 Preparation of basic medium

[0035] The formula of the basic medium of the 1L system for solid culture is: 25g glucose, 23g yeast extract, 6.25g egg yolk (other poultry egg yolks can also be substituted), egg white 6.25g (other poultry egg whites can also be substituted), chlorinated Sodium 5g, agar 2g, peanut oil 40g, add 1L distilled water to make the volume to 1L.

[0036] Prepare 1L basal medium according to the above formula, and dispense equally into 250mL conical flasks. Each 250mL conical flask is divided into 50mL basal medium.

[0037] 1.2 Preparation of nematode culture medium containing different concentrations of freeze-dried Tenebrio molitor larvae powder

[0038] Buy Tenebrio molitor larvae (available for purchase in the market), freeze for 24h and freeze-dry on a vacuum freeze dryer, dry and grind into mealworm larva powder, and store it in a refrigerator at -20°C until use.

...

Embodiment 2

[0041] Example 2: Study on the function of a medium containing freeze-dried Tenebrio molitor larvae meal

[0042] 2.1 The acquisition of entomopathogenic nematode (EPN) symbiotic bacteria

[0043] (1) Entomopathogenic nematode (EPN) symbiotic bacteria isolation and purification

[0044] EPN infects about 0.5g of wax moth (purchased from Tianjin Fitch Biotech Co., Ltd., and stored in a 14 ℃ biochemical incubator in the laboratory): prepare a 24-well plate, and place a healthy and vigorous wax moth in each hole larva. Take the Sf nematode solution to estimate its density, and ensure that 100 nematodes are added to each well and cultured at 25°C. About 30h-36h, under the ultra-clean workbench, take out the corpse of wax moth from the 24-well plate and put it in 75% alcohol for surface sterilization, cut off the second pair of gastropods of wax moth, and dip it with a disposable inoculation loop Insect lymph fluid was inoculated into NBTA identification medium (formulation: nutrient a...

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Abstract

The invention relates to the field of in-vitro solid production of nematode, in particular to a solid culture medium for nematode and a preparation method and an application thereof. The solid culturemedium for nematode includes the following components: (1) basic culture medium and (2) 1-5wt% of freeze-dried powder of Tenebrio molitor larvae, wherein the basic culture medium has no serum, and the solid culture medium for nematode does not contain lipid substances such as animal viscera and milk powder. The solid culture medium for nematode provided by the invention can significantly improvethe yield and quality of entomopathogenic nematode, and moreover, can improve the efficiency of solid production of entomopathogenic nematode, save the production time and reduce the production cost.By using nematodes produced by using the culture medium of the invention, the death of prodenia litura can be significantly accelerated, and the insecticidal efficacy of nematodes is enhanced.

Description

Technical field [0001] The present invention relates to the technical field of nematode in vitro solid production, in particular to a nematode solid culture medium and a preparation method and application thereof. Background technique [0002] With the development of agriculture, crops are easily attacked by various pests, and yields decline accordingly (I. Nyamwasa., 2018; Corrêa et al., 2016). In addition, after quantifying crop yield losses in some areas, it was found that pests have a particularly large impact, especially soil insects (Amoako-Atta, 1983; Sekamwate, 2007; Toepfer et al., 2014). After the promulgation of the new pesticide regulations, the use of illegal drugs was investigated and criminally held accountable in accordance with the law, and the safety situation of agricultural products in my country has gradually improved. However, it cannot be ignored that the phenomenon of excessive pesticide residues in vegetables in some areas is still serious (Deng Bo et al...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01K67/033
CPCA01K67/033
Inventor 阮维斌甄世玉侯艳丽李阳杜小玲
Owner NANKAI UNIV
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