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Method for culturing and separating pratylenchus

A short-bodied nematode, first nematode technology, applied in the direction of animal husbandry, etc., can solve the problems of difficulty in cleaning or separation, labor, and difficulty in removing carrot tissue residues, etc.

Active Publication Date: 2021-01-19
INST OF PLANT PROTECTION HEBEI ACAD OF AGRI & FORESTRY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Wherein, the shortcoming of existing callus culture method is: 1) the sterilization process before preparing carrot slices is more loaded down with trivial details, and what even has is also disinfected with toxic disinfectant --- mercury liter, unsafe; 2) single The number of nematode populations cultured from the piece of callus is too low, and the culture time is longer; 3) For the cultured nematodes, it is difficult to separate the nematodes located inside the callus; 3) When separating the nematodes inside the callus It is usually accompanied by a large number of dead nematodes, which greatly interferes with the follow-up experiment, and also consumes a lot of time and labor, which is not conducive to the normal operation of the experiment.
However, these methods all involve the use of a large number of mechanical methods to crush the carrot callus before separating the nematodes, which causes great damage to the nematodes, and there will be a large amount of carotene pigment and tiny carrots in the solution of the isolated nematode population. Including tissue residues, it is difficult to remove these fine carrot tissue residues even through multiple washings, and more importantly, it contains a large number of dead nematodes, which will cause difficulties in cleaning or separating the worms used in subsequent experiments, and consume a lot of time and labor

Method used

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  • Method for culturing and separating pratylenchus
  • Method for culturing and separating pratylenchus
  • Method for culturing and separating pratylenchus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043]The effect of different constant temperature conditions on the reproduction of short-body nematodes

[0044]1) Preparation of carrot callus: buy fresh red carrots on the market, first wash them with tap water, then rinse the carrots with sterile water, dry them, and then use sterile tweezers to take 75% alcohol cotton balls to wipe the surface of the carrots 1 Then put the carrots in the ultra-clean table and irradiate them with ultraviolet rays for 15 minutes. Then wipe the surface of the carrots again with a 75% alcohol cotton ball. Use a sterilized knife to cut away the turquoise base and the tip of the carrot. After sterilizing the knife with an alcohol lamp, use the knife to peel off the carrot skin, and then cut into 1.1-1.4cm thick carrot slices. Use sterilized tweezers to pick up the carrot slices and place them in a 70mm diameter sterilized petri dish. There is no need to add any culture medium inside, put 1 piece in each petri dish, seal it with a parafilm, place it in ...

Embodiment 2

[0054]The effect of low temperature pretreatment on the reproduction of Brachylenchus

[0055]In step 3), the carrot callus after the inoculation of Brachyma scribnerella in step 2) was placed in an incubator at 13°C, 15°C, 17°C, and 20°C and cultured at each temperature in the dark and constant temperature. After 4d, 6d, 8d, and 10d, they were cultured at 25℃ for 7 weeks, and then the number of reproductive nematodes of Brachymys scrimbner was investigated by dilution counting method and the reproduction coefficient was calculated. A total of 16 treatments, each treatment repeated 5 petri dishes of carrot callus.

[0056]Others are the same as in Example 1.

[0057]See Table 3 for the results of offspring reproduction, and Table 4 for the results of reproduction coefficient.

[0058]Table 3 The effects of different low-temperature pretreatments on the progeny of Brachymys scrimbner

[0059]

[0060]The data in the table is the mean ± standard error. Different lowercase letters after the same column ...

Embodiment 3

[0066]The effect of different culture time on the reproduction of short body nematodes

[0067]In step 3), the carrot callus after inoculation of Brachyteoma scribnera in step 2) is first placed in a 17°C incubator and treated at low temperature for 8 days, and then transferred to a 25°C incubator, respectively at 25°C Cultivate for 49d, 53d, 57d, 64d, 70d, 78d, 90d. A total of 7 treatments, each treatment repeated 5 petri dishes of carrot callus. The dilution counting method was used to investigate the reproduction amount of Brachymys scribneria and calculate the reproduction coefficient.

[0068]Others are the same as in Example 1.

[0069]The results are shown in Table 5.

[0070]Table 5 The effect of Brachymys scrimbner on the reproduction of nematodes after cultured at 25℃ for different days

[0071]

[0072]The data in the table is the mean ± standard error. Different lowercase letters after the same column of data indicate that they are tested by Duncan's new multiple range method in PThe diff...

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Abstract

The invention relates to a method for culturing pratylenchus and separating the pratylenchus. The method comprises the following steps of 1) inoculating pratylenchus to a culture medium; 2) treating the pratylenchus at a temperature lower than the temperature at which the pratylenchus propagates; 3) culturing the pratylenchus at a propagation temperature; and 4) harvesting the pratylenchus.

Description

Technical field[0001]The present invention relates to a method for culturing and isolation of Brachymys.Background technique[0002]Wheat root rot nematodes (Root lesion nematodes) disease is harmful all over the world. The pathogenic nematode that causes the disease is root rot nematodes, belonging to the genus Pratylenchus spp., second only to root-knot nematodes and cyst nematodes The world’s third largest nematode disease. In a large number of foreign reports on the damage of wheat crops by short-body nematodes, the main species involved include P.hexincisus, P.loosi, and P.neglectus. ), P. penetrans, P. scribneri, P. thornei, P. zeae, etc. Of the above-mentioned types of Brachyidia, except for Brachychia hexaseptide which has not been found on wheat in China, the other types have been found. In addition, there are Brachychia frugiperda, Brachyura coffea, and Brachyura agile in wheat. . The pathogenic nematode harms many crops and hosts a wide range of crops, such as wheat, oats, ...

Claims

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Application Information

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IPC IPC(8): A01K67/033
CPCA01K67/033
Inventor 李秀花马娟高波王容燕陈书龙李焦生
Owner INST OF PLANT PROTECTION HEBEI ACAD OF AGRI & FORESTRY SCI