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Cucumber downy mildew sporangium in vitro conservation method

A technology of in vitro preservation and sporangia, applied in fungi and other directions, can solve the problems of only 3.5% germination rate of sporangia, no preservation method, and easy loss of pathogenicity of sporangia.

Inactive Publication Date: 2009-05-20
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

②Preservation of isolated leaves: Fu Shuyun (1983) of Shenyang Agricultural University developed an in vitro culture method for leaves with fungi. At -5°C, diseased leaves can be preserved for 26 days, but the sporangia germination rate is only 3.5%.
④Liquid nitrogen storage: Gulya reported in 1993 that the dry sporangia of sunflower and other downy mildew fungi can be stored in liquid nitrogen without cryoprotectant, but this method requires drying and pretreatment of the sporangia. Virus, and liquid nitrogen equipment is required for storage, and stored in liquid nitrogen all year round
[0003] As can be seen from the above preservation methods about cucumber downy mildew strains, no ideal, simple and effective preservation method has been found so far.

Method used

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  • Cucumber downy mildew sporangium in vitro conservation method

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Experimental program
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Effect test

specific Embodiment approach 1

[0013] Specific implementation method one: collect fresh diseased leaves of cucumber downy mildew, wash away the sundries and original sporangia on the leaf surface, keep moist for 24 hours at 20°C to 25°C, and wait until a large number of thick and fresh sporangia grow out , using a sterilized soft brush to brush the sporangia into the mixed preservation solution with a concentration of 10% dimethyl sulfoxide and 5% skim milk in water, and then preserve it at -20°C.

specific Embodiment approach 2

[0014] Specific embodiment two: collect the fresh diseased leaves of cucumber downy mildew, wash away the sundries and original sporangia on the leaf surface, keep moist for 24 hours at 20°C to 25°C, and wait for a large amount of thick and fresh sporangia to grow , using a sterilized soft brush to brush the sporangia into the mixed preservation solution with a concentration of 10% dimethyl sulfoxide and 5% skim milk in water, and then preserve it at -70°C.

specific Embodiment approach 3

[0015] Specific implementation mode three: collect fresh diseased leaves of cucumber downy mildew, wash away the sundries and original sporangia on the leaf surface, keep moist for 24 hours at 20°C to 25°C, and wait until a large amount of thick and fresh sporangia grow out , use a sterilized soft brush to brush the sporangia into the mixed storage solution with a weight percentage concentration of 10% dimethyl sulfoxide and 5% skim milk aqueous solution, first treat it at -20°C for 24 hours, and then treat it at -80°C Store under ℃ conditions.

[0016] The steps of the above-mentioned specific implementation mode are:

[0017] 1. Source of bacteria

[0018] Collect fresh diseased leaves from cucumber downy mildew in the field, wash away the debris and original sporangia on the leaf surface, keep moist for 24 hours at 20°C to 25°C, and wait until a large number of dense and fresh sporangia grow, For the preservation of germs.

[0019] 2. Save method and steps

[0020] Brus...

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Abstract

The invention provides a in vitro conservation method for Pseudoperonospora cubensissporangium. It includes process of: the treatment and conservation method and conditions ofPseudoperonospora cubensissporangium, the detection of sporangium germination rate and pathogenicity. The invention is to conserve thePseudoperonospora cubensissporangium in a mixed liquid containing 10% dimethyl sulfoxide and 5% skimmed milk, at -20DEG C, -70DEG C, or pretreat at -20DEG C for 24h, and conserve at -70DEG C. The best conservation method is pretreating at -20DEG C for 24h, and conserving at -70DEG C, after 12 months conservation, the sporangium germination rate is 46%, disease incidence and pathogenetic condition index are 50% and 40 respectively after inoculate to cucumber seminal leaves, high pathogenicity is preserved. The invention is fit for the long term in vitro conservation of Pseudoperonospora cubensis, and can solve the problem of serious pathogenicity descent in vitro conservation fundamentally.

Description

(1) Technical field [0001] The invention relates to a method for long-term in vitro preservation of sporangia of peronospora cucumber. (2) Background technology [0002] Cucumber downy mildew is a devastating disease on cucumbers in protected fields. The disease is an airborne disease caused by the infection of Flagellate subphylum, Oomycetes, Peronosporaceae, and Pseudoperonospora cubensis (Berk. & M.A.Curtis) Rostovzev. So far, the research on the physiological differentiation and genetic diversity of the disease is still incomplete. Tracing it to its cause is mainly because Cuban pseudoperonospora is an obligate parasite, which cannot be cultivated and preserved with artificial medium. At present, the main methods of strain preservation are as follows: ① Preservation of live hosts: under suitable environmental conditions, the bacteria are regularly connected to new hosts. This is a simple but time-consuming method and requires certain equipment, but can be preserved fo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14
Inventor 张艳菊秦智伟周秀艳
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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