Method for breeding, culturing and preserving paratylenchus by using carrot callus

A callus and carrot technology, applied in the direction of botany equipment and methods, gardening methods, applications, etc., to achieve the effect of easy operation and easy acquisition

Inactive Publication Date: 2013-04-03
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The object of the present invention is to aim at the above-mentioned deficiency in the prior art of needle nematode artificial propagation and culture preservation, provide a kind of method that utilizes carrot callus propagation and culture preservation needle nematode, this method can realize simple, high-efficiency large-scale reproduction of needle nematode purpose, while preserving needle nematodes for a long time and maintaining the parasitic and pathogenicity of needle nematodes to plants

Method used

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  • Method for breeding, culturing and preserving paratylenchus by using carrot callus
  • Method for breeding, culturing and preserving paratylenchus by using carrot callus
  • Method for breeding, culturing and preserving paratylenchus by using carrot callus

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Embodiment 1

[0029] The propagation of embodiment 1 needle nematode on the carrot callus

[0030] (1) Preparation of carrot callus

[0031] Select healthy, uninjured carrots that have just been harvested within 2 weeks, wash and dry them, soak them in 95% (V / V) industrial alcohol for 2 minutes in an ultra-clean workbench, then burn them on an alcohol lamp, repeat 3 times, until the carrots The skin turns from orange red to reddish brown; peel off the carrot skin with a sterile knife, cut into thin slices about 5mm thick, and put them in a sterilized petri dish (such as figure 1 shown), sealed with plastic wrap, placed in a constant temperature incubator at 25°C for dark cultivation, and carrot callus grew in about 2 weeks for later use (such as figure 2 shown).

[0032] (2) Isolation of needle nematodes

[0033] Use the Berman funnel method to separate needle nematodes from the collected diseased roots and diseased soil. The specific method is as follows: select a glass funnel with a d...

Embodiment 2

[0038] The culture preservation of embodiment 2 needle nematodes on the carrot callus

[0039] (1) See Example 1 for the preparation of carrot callus

[0040] (2) Disinfection and inoculation of needle nematodes

[0041] When the yellow-brown or dark-brown lesions appear on the carrot callus of the cultured and propagated nematodes in the early stage, in the ultra-clean workbench, wash the nematodes in the culture dish with sterile water and suck them into the centrifuge tube, and then press In the disinfection and inoculation method described in Example 1, the needle nematodes were sterilized and inoculated onto the freshly prepared carrot callus, placed in a constant temperature incubator, and cultured and stored in the dark at 23°C. After 16 weeks, most of the callus of carrot beetroot showed yellow-brown or dark-brown lesions and live needle nematodes existed.

[0042] (3) Subculture of needle nematodes

[0043] Yellow-brown lesions appear in the carrot callus part of t...

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Abstract

The invention discloses a method for breeding, culturing and preserving paratylenchus by using carrot callus, belonging to the field of plant-parasitic nematode culture. The method comprises the following steps of: (1) under a sterile condition, soaking carrots with alcohol, scorching, peeling and slicing the carrots, putting carrot slices into a sterilized culture dish, sealing an opening of the sterilized culture dish, putting into a culture box, and culturing till the carrot slices grow callus; (2) sterilizing the paratylenchus with a streptomycin sulfate solution, centrifuging and collecting, eluting with sterile water, centrifuging and collecting again, preparing a paratylenchus suspension with sterile water, and dripping the suspension on the carrotcallus; and (3) putting the carrot callus inoculated with the paratylenchus into the culture box, and culturing. In the method, a culture medium is not required for culturing, so that the operation is simple and convenient; and sterilization of the streptomycin sulfate solution does not affect the activity of the paratylenchus, so that the aim of breading the paratylenchus simply, efficiently and largely is achieved, the paratylenchus can be preserved for a long time, and parasitism and pathogenicity of the paratylenchus on a plant are kept.

Description

technical field [0001] The invention relates to an artificial culture method for propagating and preserving plant parasitic nematodes by using plant callus, in particular to a method for propagating, cultivating and preserving needle nematodes by using carrot callus. Background technique [0002] Needle nematode (Paratylenchus spp.) belongs to Tylenchida, Tylenchina, Criconematoidea, Tylenchulidae, Paratylenchinae, is a kind of ectoparasitic plant nematode, mainly Infect the roots and other underground tissues of parasitic plants, affecting the normal growth of plants, resulting in a decline in yield and quality. To measure and systematically study the biology, pathology, ecology and control methods of needle nematodes, a large number of purified individuals are required and stored for a long time, which requires artificial propagation and cultivation of needle nematodes. There have been studies on the use of white clover (Trifolium repens) callus to culture Paratylenchus p...

Claims

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Application Information

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IPC IPC(8): A01H4/00A01K67/033
Inventor 谢辉王珂徐春玲李宇
Owner SOUTH CHINA AGRI UNIV
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