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Method for separating living eggs of meloidogyne incognita

A technology of M. incognita and an isolation method, which is applied in the field of agricultural plant protection, can solve the problems of inability to separate oocysts, inability to accurately assess the number of M. incognita, inability to guarantee live eggs and the like, and achieve the effect of ensuring activity

Inactive Publication Date: 2021-02-09
陕西省生物农业研究所
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] There are many separation methods for root-knot nematodes, including Berman funnel method, shallow dish method, centrifugation method, centrifugal floating method and elutriation and dyeing method. These methods have their own advantages and disadvantages. 2nd instar nematode larvae active in the soil, but eggs cannot be separated; centrifugation, centrifugal floatation and elutriation and staining methods can only separate nematode larvae and scattered eggs in the soil, but cannot separate oocysts attached to the root system, and are often different Eggs of different types of nematodes are mixed, and there is no guarantee that the eggs are alive
However, root-knot nematodes mostly attach to the root surface or soil particles in the form of oocysts to survive the unfavorable environment, so traditional methods cannot accurately assess the number of root-knot nematodes

Method used

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  • Method for separating living eggs of meloidogyne incognita
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  • Method for separating living eggs of meloidogyne incognita

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: Effects of Different Cultivation Time of Meloidogyne incognita oocysts on Oocyst Dissociation

[0029] 1) Materials and methods: collect diseased roots of pumpkins infested by root-knot nematode incognita, and keep them moist at 28°C; after 12, 24, 48, and 72 hours, cut a small amount of root tissue and rinse thoroughly with tap water; Under the stereoscope, pick 10 (milky white) M. incognita oocysts of the same size with tweezers from the diseased roots processed at different times respectively, wherein each processing comprises 5 independent technical repetitions (replicate); The picked nematode oocysts were placed in a 1.5 mL centrifuge tube containing 500 μL of sterile water; after centrifugation at 1600 rpm for 2 min, the supernatant was fully removed with a pipette; 1 mL of sodium hypochlorite (NaClO) with an effective concentration of 0.3% was added ) solution to treat nematode oocysts for 2 minutes, during which the samples were repeatedly shaken on a...

Embodiment 2

[0031] Example 2 Effects of Different NaClO Treatment Concentrations on the Recovery and Hatching of Meloidogyne incognita Eggs

[0032] 1) Materials and methods: collect diseased roots of pumpkins infested by root-knot nematode incognita, and place them at 28°C (moisturizing) for 48 hours; after that, rinse the diseased root tissue thoroughly with tap water; under a stereoscope, pick 10 seeds of the same size with tweezers. One (milky white) root-knot nematode oocyst; put the picked nematode oocyst into a 1.5mL centrifuge tube containing 500μL sterile water; centrifuge at 1600rpm for 2min, and remove the supernatant with a pipette , divided into 5 parts; in each centrifuge tube containing oocysts, add 1 mL of sodium hypochlorite (NaClO) solution with an effective concentration of 0% (sterile water), 0.3%, 0.5%, 1.0%, 1.5% and 2% respectively , wherein each treatment contained 5 independent technical repetitions (replicate); the sample was shaken repeatedly on a vortex shaker,...

Embodiment 3

[0036] Embodiment 3: The influence of different treatment time of NaClO on the egg hatching rate of root-knot nematode incognita

[0037]1) Materials and methods: Plant diseased roots placed at 28°C (moisturizing) for 48 hours were thoroughly rinsed with tap water, and 10 (milky white) M. incognita oocysts were picked out with tweezers; In a 1.5mL centrifuge tube containing 500μL sterile water; after centrifugation at 1600rpm for 2min, remove the supernatant fully with a pipette; in each centrifuge tube containing oocysts, add 1mL with an effective concentration of 0.3% sodium hypochlorite ( NaClO) solution was processed sample 2, 5, 8 and 10min respectively, during which the sample was shaken repeatedly on the vortex shaker, wherein each processing contained 5 independent technical repetitions (replicate); The sample was centrifuged 2min under the condition of 1600rpm, Discard the supernatant, and fully wash the sample with sterile water for 3 times; add 500 μL of sterile wat...

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Abstract

The invention discloses a method for separating living eggs of meloidogyne incognita. The method comprises the following steps of stripping meloidogyne incognita oocysts attached to the surface of a root system to be subjected to static incubation, NaOCl dissociation, cleaning, oscillation, centrifugation and the like, so that pure meloidogyne incognita eggs with a dispersed single population canbe separated, meanwhile, the activity of the eggs is ensured, and value for popularization and application is achieved.

Description

technical field [0001] The invention relates to the field of agricultural plant protection, in particular to a method for separating live eggs of root-knot nematode incognita. Background technique [0002] Root-knot nematode eggs are gathered together and exist in gelatin coated with egg masses. During the experiment, it is often necessary to study the hatching, development and biological characteristics of root-knot nematode eggs and second-instar larvae, which is an important experimental application material. , but it is difficult to separate the collected eggs into individual eggs. This technology mainly separates the eggs of Meloidogyne incognita efficiently to obtain high-purity and impurity-free nematode eggs and ensure their activity to solve this problem. Applied materials are provided during relevant experimental studies. [0003] Root-knot nematode disease is mainly caused by root-knot nematode (Meloidogyne incognita) infestation, and its occurrence range include...

Claims

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Application Information

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IPC IPC(8): A01K67/033
CPCA01K67/033
Inventor 张锋王远征张淑莲李英梅洪波杨艺伟付博任平
Owner 陕西省生物农业研究所
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