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Insect tissue zymolyte and use thereof in artificial culture of entomopathogenic nematodes

An entomopathogenic nematode and artificial culture technology is applied in the field of entomopathogenic nematode culture medium containing enzymatic hydrolysates of insect tissues, which can solve the problems of increasing the difficulty of cleaning nematodes and reducing the cleanliness of nematodes, and achieves short culture cycle, low production cost, Easy to operate effect

Inactive Publication Date: 2009-04-22
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the presence of insect cuticles in the culture medium will greatly reduce the cleanliness of nematodes and increase the difficulty of cleaning nematodes when they are harvested

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Contrast experiment on culture of Steinernema feltiae

[0026] 1. Preparation of enzymatic hydrolyzate of insect Mellonella mellonella

[0027] (1) Select 16 g of the last instar larvae of the greater wax moth moth with the same nutritional and physiological conditions cultivated in the laboratory, add 500 g of water, and use a tissue homogenizer to fully break the insect tissue, so that the tissue contents of the greater wax moth are completely released and evenly suspended in the in the mixture;

[0028] (2) Add 1 g of neutral protease (Beijing Aoboxing Company) to the mixture, and carry out hydrolysis reaction at 45° C. for 10 minutes; then use dense silk cloth to filter out the residue of the wax moth tissue, and the obtained filtrate is the tissue of the wax moth Enzymolyzate.

[0029] 2. Preparation of culture medium for Steineria turnipii

[0030] (1) Add 240g of soybean powder, 80g of flour, 16g of egg yolk powder, 8g of peptone, 24g of yeast extract, 80g of ...

Embodiment 2

[0041] Embodiment 2 Contrast experiment on the culture of Steineria turnipii

[0042] Repeat Example 1, but make the following modifications: Change 1 g of neutral protease in Step 1 (2) of Example 1 to 1 g of acid protease (Beijing Aoboxing Company), and change the enzymatic hydrolysis reaction conditions to hydrolysis at 55 ° C React for 30 minutes.

[0043] The results (see Table 2) yield of nematodes in insect tissue culture was 1.50 times that of bean flour medium, the culture period was shortened by 6 days compared with bean flour medium, and the invasion rate of harvested nematodes in the infestation period to Greater Melonella mellonella increased by 6.10%.

[0044]Table 2 Results of comparative experiments on culture of Steineria turnipii

[0045] culture medium Yield (IJs / g) Training cycle nematode invasion rate soybean flour medium 2.53×10 5 22 days 3.22％ Insect tissue culture medium 3.80×10 5 16 days 9.32％

Embodiment 3

[0046] Embodiment 3 Contrast experiment on the culture of Steineria turnipii

[0047] Repeat Example 1, but make the following modifications: Change the neutral protease in Example 1 step 1 (2) to 0.5 g of alkaline protease (Beijing Aoboxing Company), and the enzymolysis reaction condition is to carry out hydrolysis at 50 ° C React for 60 minutes;

[0048] Result (see table 3) insect tissue culture nematode output is 1.58 times of contrast bean flour medium, and culture cycle shortens 6 days than contrast bean flour medium, and the invasion rate of nematode in the infestation period of harvest has improved 4.78 %.

[0049] Table 3 The results of the comparison test of the culture of Steineria turnipii

[0050] culture medium Yield (IJs / g) Training cycle nematode invasion rate soybean flour medium 2.53×10 5 22 days 3.22％ Insect tissue culture medium 4.01×10 5 16 days 8.00％

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Abstract

The invention discloses an insect tissue zymolyte. The protease is used for hydrolyzing the insect tissue which is then filtrated to obtain filter liquor; and the filter liquor is the insect tissue zymolyte. The invention also discloses the application of the insect tissue zymolyte on the artificial culture of entomopathogenic nematodes. The insect tissue zymolyte is added into a conventional culture medium as a component, and then is blended evenly and sterilized. The nematodes cultured in the culture medium prepared by the insect tissue zymolyte have high yield, a short culturing cycle, strong virulence, and low cost. The nematodes can be harvested more cleanly, thereby simplifying the harvesting procedure and reducing the probability of pollution during the nematodes storage period. The insect tissue zymolyte has simple and feasible operation, and is suitable for large-scale production.

Description

technical field [0001] The invention belongs to the field of biological control of plant pests, and in particular relates to an enzymatic insect tissue hydrolyzate, the application of the enzymatic insect tissue hydrolyzate in the artificial cultivation of entomopathogenic nematodes, and the cultivation of entomopathogenic nematodes containing the enzymatic insect tissue hydrolyzate base. Background technique [0002] Entomopathogenic nematodes are an important class of biological insecticides. They enter the insect body along with the food of the host insect or from the insect's natural opening and intersegmental membrane, and release the symbiotic bacteria it carries in the insect blood cavity, and the virulence factors secreted by the symbiotic bacteria Cause insects to suffer from septicemia and die. Entomopathogenic nematodes have a wide range of hosts and can actively search for hosts. Therefore, they have unique control advantages against hidden pests such as soil-dw...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A23K1/18A01K67/033A23K10/14A23K50/00
Inventor 简恒曹翠玲刘倩王金利
Owner CHINA AGRI UNIV
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