Separation method of meloidogyne incognita eggs

A technology for the isolation of root-knot nematode incognita, applied in the field of agricultural plant protection, can solve the problems of research, inability to realize well, unfavorable biological characteristics of nematode eggs, etc., and achieve the effect of high egg activity

Pending Publication Date: 2022-03-15
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, such as Berman funnel separation method, shallow plate separation method, floating-sieving separation method, sucrose separation method and semi-automatic elutriation method, etc. These methods have their own advantages and disadvantages, and the separation Live (mixed) nematode populations are the main purpose, but the separation of eggs cannot be well achieved
For direct anatomy, pulverization and screening, and high-concentration sodium hypochlorite digestion, although a small amount of nematode eggs can be separated, the purity and vigor of the obtained eggs are significantly affected, which is not conducive to the extraction of nematode eggs. Research on biological characteristics

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  • Separation method of meloidogyne incognita eggs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1: Effects of different lengths of tending on egg mass dissociation of Meloidogyne incognita

[0028] 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 a stereoscope, pick 10 (milky white) egg masses of M. incognita with tweezers from the diseased roots treated at different times, each of which contains 5 independent technical replicates (replicate); will pick The nematode egg masses were placed in a 1.5mL centrifuge tube containing 500μL sterile water; after centrifugation at 1600rpm for 2min, the supernatant was fully removed with a pipette; 1mL of sodium hypochlorite (NaClO) solution with an effective concentration of 0.3% was added to treat the nematodes Egg mass for 2 minutes, during which the samples were shaken repeatedly on a vortex shaker; centrifuge at 1600rpm for ...

Embodiment 2

[0030] Example 2 Effects of Different NaClO Concentrations on Meloidogyne incognita Egg Dissociation and Egg Hatching

[0031] 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) egg mass of Meloidogyne incognita; put the picked nematode egg mass into a 1.5mL centrifuge tube containing 500μL sterile water; centrifuge at 1600rpm for 2min, remove the supernatant with a pipette and divide into 5 parts ; In each centrifuge tube containing egg pieces, 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 Including 5 independent technical repetitions (replicate); the sample was shaken repeatedly on a vortex shaker...

Embodiment 3

[0035] Embodiment 3: The influence of different treatment time of NaClO on hatching of eggs of Meloidogyne incognita

[0036] 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) egg masses of M. incognita were picked out with tweezers; into 1.5mL centrifuge tubes containing bacterial water; after centrifugation at 1600rpm for 2min, remove the supernatant fully with a pipette; add 1mL of 0.3% sodium hypochlorite (NaClO) solution with an effective concentration of 0.3% to each centrifuge tube containing egg masses for treatment Samples 2, 5, 8 and 10min, during which the samples were repeatedly shaken on a vortex shaker, wherein each treatment contained 5 independent technical repetitions (replicate); the samples were centrifuged at 1600rpm for 2min, and the supernatant was discarded. Wash the samples 3 times with sterile water; add 500 μL sterile water, mix the samples and let them...

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Abstract

The invention discloses a method for separating meloidogyne incognita egg granules, which comprises the following steps of: performing static incubation, NaClO dissociation, oscillation, centrifugation, cleaning and the like on meloidogyne incognita egg blocks stripped from the surface of a plant root system, so that pure meloidogyne incognita egg granules can be separated, meanwhile, higher egg activity is ensured, and the method has popularization and application values.

Description

technical field [0001] The invention relates to the field of agricultural plant protection, in particular to a method for separating eggs of root-knot nematode incognita. Background technique [0002] The root-knot nematode Meloidogyne incognita can obligately parasitize the root system of plants. While affecting the normal morphology of root tissue, destroying nutrition and water absorption and metabolism, it can also lead to inhibition of plant nutrition and reproductive growth, such as short plants, wilting of yellow leaves, and fruit setting. rate reduction etc. Meloidogyne incognita, because of its short life cycle, wide range of hosts, strong environmental adaptability, and the combination of plant pathogenic bacteria and fungi, causes economic losses of up to 50 billion yuan in the development of my country's vegetable industry every year. More importantly, with the scale and intensification transformation and upgrading of my country's vegetable and fruit industry, c...

Claims

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Application Information

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IPC IPC(8): A01K67/033
CPCA01K67/033
Inventor 王远征
Owner 陕西省生物农业研究所
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