CN103645191A - Method for realizing rapid detection of phenomenon of identifying injurious insect host plant roots by entomopathogenic nematodes - Google Patents
Method for realizing rapid detection of phenomenon of identifying injurious insect host plant roots by entomopathogenic nematodes Download PDFInfo
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- CN103645191A CN103645191A CN201310653377.2A CN201310653377A CN103645191A CN 103645191 A CN103645191 A CN 103645191A CN 201310653377 A CN201310653377 A CN 201310653377A CN 103645191 A CN103645191 A CN 103645191A
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Abstract
The invention provides a method for realizing rapid detection of a phenomenon of identifying injurious insect host plant roots by entomopathogenic nematodes. The invention relates to a simple, direct and novel efficient detection method for a plant root signal reaction by the entomopathogenic nematodes. The method comprises the following steps: 1, preparing pre-cooling sterilizing distilled water; 2, preparing Pluronic liquid-state glue; 3, putting the entomopathogenic nematodes at an infection period into the Pluronic liquid-state glue; uniformly agitating and transferring a mixture into a plastic vessel; then putting plant seedling roots and standing for a period of time at 20-30 DEG C; and observing by an anatomical lens to realize the rapid detection of the phenomenon of identifying the injurious insect host plant roots by the entomopathogenic nematodes. The method is mainly used for observing the phenomenon of identifying the injurious insect host plant roots by the entomopathogenic nematodes.
Description
Technical field
The present invention relates to simple, directly perceived, the novel efficient detection method of a kind of entomopathogenic nematode to plant roots signal reaction.
Background technology
Entomopathogenic nematode (EntomopathogenicNematode, EPN) is the Important Natural Enemy of insect, has host insect widely, is the important means of current biological control of insect pests.The preventive and therapeutic effect of entomopathogenic nematode to insect, the parasitic effects of the entomopathogenic nematode of the indoor soil bioassay research of most employings in the past to insect, and potted plant and prevention effect field, but entomopathogenic nematode is different to the prevention effect of the subterranean pest-insect of the same race of crop of the same race not, illustrates that crop root environment not of the same race has a certain impact to entomopathogenic nematode.In default of detection method accurately, the effect of crop root microenvironment not of the same race in controlling underground pest is never elaborated.
Summary of the invention
The invention provides a kind of method of fast detecting entomopathogenic nematode to insect host plant root identification phenomenon that realize.
A kind of method of fast detecting entomopathogenic nematode to insect host plant root identification phenomenon that realize, specifically complete according to the following steps: one, first distilled water is carried out to moist heat sterilization processing, then in temperature, be precooling 1h～5h at 4～10 ℃, obtain precooling sterile purified water; Two, Pluronic rubber powder being poured in precooling sterile purified water, is then to stir 16h～32h at 4～10 ℃ in temperature, obtains Pluronic liquid glue; Three, by every milliliter of Pluronic liquid glue, containing 250～350 concentration that infect phase entomopathogenic nematode, will infect phase entomopathogenic nematode and put into Pluronic liquid glue, after stirring and evenly mixing, obtain nematode glue suspension; Plastic ware is placed on ice, then nematode glue suspension is transferred in plastic ware, again by under insect host plant seedling undercut, obtain the Young Plant shoot root of 0.5cm～2cm, again the Young Plant shoot root of 0.5cm～2cm is placed in the plastic ware containing nematode glue suspension, obtain plastic ware to be seen, it is to place 20min～40min at 20～30 ℃ that plastic ware to be seen is placed in to temperature, then under anatomical lens observation of plant seedling around or on root table and cutter cut wound nematode population, realize fast detecting entomopathogenic nematode to insect host plant root identification phenomenon.
Advantage of the present invention: the present invention utilize a kind of can simulated soil physical environment, the nematode transparent adhesive tape (PluronicF-127 that can move freely therein, trade name is pluronic, principal ingredient is the addition polymer of polypropylene glycol and oxirane) system, provide accurately, fast, detection method intuitively, resolved the effect of crop root microenvironment not of the same race in entomopathogenic nematode control subterranean pest-insect.Pluronic glue, being in a liquid state lower than 15 ℃, is solid-state when higher than 15 ℃, but nematode can be movable freely in this solid-state glue, and Pluronic glue is in being usually used in cosmetics, to nematode nonhazardous effect.
Embodiment
Embodiment one: present embodiment is a kind of method of fast detecting entomopathogenic nematode to insect host plant root identification phenomenon that realize, specifically complete according to the following steps: one, first distilled water is carried out to moist heat sterilization processing, then in temperature, be precooling 1h～5h at 4～10 ℃, obtain precooling sterile purified water; Two, Pluronic rubber powder being poured in precooling sterile purified water, is then to stir 16h～32h at 4～10 ℃ in temperature, obtains Pluronic liquid glue; Three, by every milliliter of Pluronic liquid glue, containing 250～350 concentration that infect phase entomopathogenic nematode, will infect phase entomopathogenic nematode and put into Pluronic liquid glue, after stirring and evenly mixing, obtain nematode glue suspension; Plastic ware is placed on ice, then nematode glue suspension is transferred in plastic ware, again by under insect host plant seedling undercut, obtain the Young Plant shoot root of 0.5cm～2cm, again the Young Plant shoot root of 0.5cm～2cm is placed in the plastic ware containing nematode glue suspension, obtain plastic ware to be seen, it is to place 20min～40min at 20～30 ℃ that plastic ware to be seen is placed in to temperature, then under anatomical lens observation of plant seedling around or on root table and cutter cut wound nematode population, realize fast detecting entomopathogenic nematode to insect host plant root identification phenomenon.
Pluronic glue, being in a liquid state lower than 15 ℃, is solid-state when higher than 15 ℃, but nematode can be movable freely in this solid-state glue, and Pluronic glue is in being usually used in cosmetics, to nematode nonhazardous effect.The solid-state glue of Pluronic that present embodiment utilizes step 2 to prepare replaces soil, because when Pluronic liquid glue is to become the solid-state glue of Pluronic at 20～30 ℃ in temperature, and transparent during the solid-state glue of Pluronic, therefore utilize anatomical lens can observe direction and speed that nematode is moved in Pluronic liquid glue, realize fast detecting entomopathogenic nematode to insect host plant root identification phenomenon, and cut off according to counting plant roots, the nematode population of assembling on wound and root surface, analyze and judge that kindred plant root is not to the attraction of entomopathogenic nematode or repulsive interaction.
Embodiment two: the difference of present embodiment and embodiment one is: it is as follows that moist heat sterilization described in step 1 is processed concrete operations: be sterilization treatment 30min under 121 ℃ and 1 atmospheric pressure in temperature.Other are identical with embodiment one.
Embodiment three: present embodiment and one of embodiment one or two difference are: first distilled water being carried out to sterilization treatment in step 1, is then precooling 1h～3h at 4～10 ℃ in temperature, obtains precooling sterile purified water.Other are identical with embodiment one or two.
Embodiment four: one of present embodiment and embodiment one to three difference is: in step 2, Pluronic rubber powder being poured in precooling sterile purified water, is then to stir 24h at 4～10 ℃ in temperature, obtains Pluronic liquid glue.Other are identical with embodiment one to three.
Embodiment five: one of present embodiment and embodiment one to four difference is: will infect phase entomopathogenic nematode by every milliliter of Pluronic liquid glue containing 300 concentration that infect phase entomopathogenic nematode in step 3 and put into Pluronic liquid glue, and obtain nematode glue suspension after stirring and evenly mixing; Then nematode glue suspension is transferred in plastic ware, plastic ware is placed on ice, then by under insect host plant seedling undercut, obtain the Young Plant shoot root of 1cm, again the Young Plant shoot root of 1cm is placed in the plastic ware containing nematode glue suspension, obtain plastic ware to be seen, it is to place 30min at 25 ℃ that plastic ware to be seen is placed in to temperature, then under anatomical lens observation of plant seedling around or on root table and cutter cut wound nematode population, realize fast detecting entomopathogenic nematode to insect host plant root identification phenomenon.Other are identical with embodiment one to four.
Content of the present invention is not limited only to the content of the respective embodiments described above, and the combination of one of them or several embodiments equally also can realize the object of invention.
Adopt following verification experimental verification effect of the present invention:
Test one: a kind of method of fast detecting entomopathogenic nematode to insect host plant root identification phenomenon that realize, specifically complete according to the following steps: one, first distilled water is carried out to sterilization treatment, then in temperature, be precooling 2h at 5 ℃, obtain precooling sterile purified water; Two, the Pluronic rubber powder of 23g being poured in 80mL precooling sterile purified water, is then to stir 24h at 5 ℃ in temperature, obtains Pluronic liquid glue; Three, by every milliliter of Pluronic liquid glue, containing 300 concentration that infect phase entomopathogenic nematode, will infect phase entomopathogenic nematode and put into Pluronic liquid glue, after stirring and evenly mixing, obtain nematode glue suspension; Then nematode glue suspension is transferred in plastic ware, plastic ware is placed on ice, then by under insect host plant seedling undercut, obtain the Young Plant shoot root of 1cm, again the Young Plant shoot root of 1cm is placed in the plastic ware containing nematode glue suspension, obtain plastic ware to be seen, it is to place 30min at 25 ℃ that plastic ware to be seen is placed in to temperature, then under anatomical lens observation of plant seedling around or on root table and cutter cut wound nematode population, realize fast detecting entomopathogenic nematode to insect host plant root identification phenomenon.
The described Young Plant shoot root of this test is respectively the plantation Radix Folium Allii tuberosi of 6 years, transplants fibrous root and the sowing tomato main root of 50 days and the fibrous root of the fibrous root point of interface 0～0.5cm of place of garlic root, the plantation shallot root of 1 year, the sowing soybean main root of 50 days and the fibrous root point of interface 0～0.5cm of place of 20 days; Above-mentioned Young Plant shoot root, by vertically cutting 1cm Young Plant shoot root from the 2.0-2.5cm of foundation portion, is to the Young Plant shoot root of 1cm.
The described phase of the infecting entomopathogenic nematode of this test is had a liking for bacterium heterorhabditis indica Heterorhabditis bacteriophora-HBN for the phase of infecting.
Utilize anatomical lens to observe every 2 hours, found that Radix Folium Allii tuberosi is the strongest to the signal of this nematode, especially in the wound that cuts off root, assemble nematode maximum, during 30min, 50-80 bar nematode is assembled in wound, two ends, next is green onion and garlic (20 left and right), soybean is the most weak to the signal of this nematode, and during 30min, assemble less than 3-8 bar nematode wound, two ends, tomato root take second place (10-14 bar).Former studies shows that entomopathogenic nematode almost reaches 100% to the prevention effect of leek root maggot, effect is better than soybean subterranean pest-insect, so infer that the root exudates that Radix Folium Allii tuberosi discharges has very strong sucking action to entomopathogenic nematode, the probability that has so greatly increased near the insect-root maggot of this nematode infection Radix Folium Allii tuberosi, causes this nematode almost to reach 100% to the prevention effect of leek root maggot.
Claims (5)
1. realize the method for fast detecting entomopathogenic nematode to insect host plant root identification phenomenon for one kind, it is characterized in that realizing fast detecting entomopathogenic nematode completes according to the following steps to the method for insect host plant root identification phenomenon: one, first distilled water is carried out to moist heat sterilization processing, then in temperature, be precooling 1h～5h at 4～10 ℃, obtain precooling sterile purified water; Two, Pluronic rubber powder being poured in precooling sterile purified water, is then to stir 16h～32h at 4～10 ℃ in temperature, obtains Pluronic liquid glue; Three, by every milliliter of Pluronic liquid glue, containing 250～350 concentration that infect phase entomopathogenic nematode, will infect phase entomopathogenic nematode and put into Pluronic liquid glue, after stirring and evenly mixing, obtain nematode glue suspension; Plastic ware is placed on ice, then nematode glue suspension is transferred in plastic ware, again by under insect host plant seedling undercut, obtain the Young Plant shoot root of 0.5cm～2cm, again the Young Plant shoot root of 0.5cm～2cm is placed in the plastic ware containing nematode glue suspension, obtain plastic ware to be seen, it is to place 20min～40min at 20～30 ℃ that plastic ware to be seen is placed in to temperature, then under anatomical lens observation of plant seedling around or on root table and cutter cut wound nematode population, realize fast detecting entomopathogenic nematode to insect host plant root identification phenomenon.
2. a kind of method of fast detecting entomopathogenic nematode to insect host plant root identification phenomenon that realize according to claim 1, it is as follows that the moist heat sterilization described in step 1 is processed concrete operations: in temperature, be sterilization treatment 30min under 121 ℃ and 1 atmospheric pressure.
3. a kind of fast detecting entomopathogenic nematode of realizing according to claim 1 is identified the method for phenomenon to insect host plant root, it is characterized in that first distilled water being carried out to sterilization treatment in step 1, then in temperature, be precooling 1h～3h at 4～10 ℃, obtain precooling sterile purified water.
4. a kind of fast detecting entomopathogenic nematode of realizing according to claim 1 is identified the method for phenomenon to insect host plant root, it is characterized in that in step 2, Pluronic rubber powder being poured in precooling sterile purified water, then in temperature, be to stir 24h at 4～10 ℃, obtain Pluronic liquid glue.
5. a kind of fast detecting entomopathogenic nematode of realizing according to claim 1 is identified the method for phenomenon to insect host plant root, it is characterized in that in step 3 that by every milliliter of Pluronic liquid glue, containing 300 concentration that infect phase entomopathogenic nematode, will infect phase entomopathogenic nematode puts into Pluronic liquid glue, obtains nematode glue suspension after stirring and evenly mixing; Then nematode glue suspension is transferred in plastic ware, plastic ware is placed on ice, then by under insect host plant seedling undercut, obtain the Young Plant shoot root of 1cm, again the Young Plant shoot root of 1cm is placed in the plastic ware containing nematode glue suspension, obtain plastic ware to be seen, it is to place 30min at 25 ℃ that plastic ware to be seen is placed in to temperature, then under anatomical lens observation of plant seedling around or on root table and cutter cut wound nematode population, realize fast detecting entomopathogenic nematode to insect host plant root identification phenomenon.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104483447A (en) * | 2014-11-26 | 2015-04-01 | 河北省农林科学院植物保护研究所 | Method for screening nematicide by utilizing pluronic F127 and sweet potato stems |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104483447A (en) * | 2014-11-26 | 2015-04-01 | 河北省农林科学院植物保护研究所 | Method for screening nematicide by utilizing pluronic F127 and sweet potato stems |
| CN104483447B (en) * | 2014-11-26 | 2016-08-24 | 河北省农林科学院植物保护研究所 | A kind of method utilizing pluronic F127 and Sweet Potato screening nematicide |
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Application publication date: 20140319 |