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A method for cultivating and isolating short-bodied nematodes

A short-bodied nematode and first nematode technology, applied in animal husbandry and other directions, can solve the problems of cumbersome disinfection process, low number of nematodes, time-consuming and other problems, and achieve the effects of improving reproduction rate, shortening culture time, and saving labor and time.

Active Publication Date: 2022-03-08
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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  • A method for cultivating and isolating short-bodied nematodes
  • A method for cultivating and isolating short-bodied nematodes
  • A method for cultivating and isolating short-bodied nematodes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Effects of different constant temperature conditions on the reproduction of Brachybody nematodes

[0044]1) Preparation of carrot callus: buy fresh red-heart carrots on the market, wash them with tap water first, then rinse the carrots with sterile water, dry them, and then use sterilized tweezers to pick up 75% alcohol cotton balls and wipe the surface of the carrots for 1 Then put the carrots in the ultra-clean table, after irradiating with ultraviolet rays for 15 minutes, then wipe the surface of the carrots again with 75% alcohol cotton balls, cut off the turquoise base end of the carrots and the tip of the carrots with a sterilized knife, After sterilizing the knife with the flame of an alcohol lamp, use the knife to peel off the skin of the carrot, then cut it into carrot slices with a thickness of 1.1-1.4cm, and use sterilized forceps 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, ...

Embodiment 2

[0054] Effects of low temperature pretreatment on reproduction of Brachybody nematodes

[0055] In step 3), the carrot calli inoculated with Brachybulina scribneri in step 2) were placed in incubators at 13°C, 15°C, 17°C, and 20°C respectively, and cultured in the dark at each temperature After 4d, 6d, 8d, and 10d, transfer to 25°C and culture for 7 weeks, then use the dilution counting method to investigate the number of reproductive nematodes and calculate the reproductive coefficient. There were 16 treatments in total, and each treatment replicated 5 culture dishes of carrot callus.

[0056] Others are the same as embodiment 1.

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

[0058] Table 3 Effects of different low-temperature pretreatments on the reproductive offspring of Brachybody scribnera

[0059]

[0060] The data in the table are mean ± standard error. Different lower...

Embodiment 3

[0066] Effects of Different Culture Time on Brevibody Nematode Reproduction

[0067] In step 3), the carrot callus after being inoculated with Brachybulina scribneri in step 2) was first placed in a 17°C incubator for low-temperature treatment for 8 days, and then transferred to a 25°C incubator, and respectively Cultured for 49d, 53d, 57d, 64d, 70d, 78d, 90d. There were 7 treatments in total, and each treatment replicated 5 culture dishes of carrot callus. The reproduction number of Brachybody scribnera was investigated by dilution counting method, and the reproduction coefficient was calculated.

[0068] Others are the same as embodiment 1.

[0069] The results are shown in Table 5.

[0070] Table 5 Effects of Brachybody scribner on nematode reproduction after culturing at 25°C for different days

[0071]

[0072] The data in the table are mean ± standard error. Different lowercase letters after the data in the same column indicate significant difference at P<0.05 le...

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Abstract

The present invention relates to a method for cultivating and isolating short-bodied nematodes, which comprises the following steps: 1) inoculating the nematodes on a culture substrate; 2) treating the nematodes at a temperature lower than the reproduction temperature of the nematodes; 3) Cultivating the nematodes at a breeding temperature; 4) Harvesting the nematodes.

Description

technical field [0001] The invention relates to a method for cultivating and isolating Brachybody nematodes. Background technique [0002] Wheat root rot nematode (Root lesion nematodes) disease is harmful all over the world. The pathogenic nematode causing the disease is root rot nematode, which belongs to the genus Pratylenchus spp., and is second only to root-knot nematodes and cyst nematodes. 3rd largest nematode disease in the world. In a large number of foreign reports on the harm of short-bodied nematodes to wheat crops, the main species involved include P.hexincisus, P.loosi, P.neglectus ), P. penetrans, P. scribneri, P. thornei, and P. zeae. For the above-mentioned short-bodied nematodes, in China, except for the six-fisted short-bodied nematodes, which have not been found on wheat, other types have been found. In addition, short-bodied nematodes grassland, coffee short-bodied nematodes, and agility short-bodied nematodes have also been found on wheat. . The pat...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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