CN106962324A - A kind of method of long-term storage entomopathogenic nematode - Google Patents
A kind of method of long-term storage entomopathogenic nematode Download PDFInfo
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- CN106962324A CN106962324A CN201710297024.1A CN201710297024A CN106962324A CN 106962324 A CN106962324 A CN 106962324A CN 201710297024 A CN201710297024 A CN 201710297024A CN 106962324 A CN106962324 A CN 106962324A
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- nematode
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/02—Preservation of living parts
- A01N1/0205—Chemical aspects
- A01N1/021—Preservation or perfusion media, liquids, solids or gases used in the preservation of cells, tissue, organs or bodily fluids
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F11/00—Other organic fertilisers
- C05F11/02—Other organic fertilisers from peat, brown coal, and similar vegetable deposits
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
- C05G3/60—Biocides or preservatives, e.g. disinfectants, pesticides or herbicides; Pest repellants or attractants
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- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Pest Control & Pesticides (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Plant Pathology (AREA)
- Dentistry (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
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- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention discloses a kind of method of long-term storage entomopathogenic nematode.Comprise the following steps：By nematode mud and fertile soil according to mass ratio 1:0.8~1.2 is well mixed, and then mixture is dispensed into container, stored after being sealed with breathable sealing film；Described nematode mud is made up of entomopathogenic nematode infection period nematode and water.The method of the present invention have the advantages that raw material be easy to get, easily operation, cost is low, occupy little space, period of storage length, survival rate height etc..The nematodes of S.longicaudum X 7 press 1 with fertile soil:0.8~1:1.2 mixtures are up to 180d after 5 DEG C of stable storages, and survival rate is still greater than 90%.The entomopathogenic nematode of storage, which can be directly spread fertilizer over the fields to field, to be used to prevent and treat Field Pests, while field fertility can also be improved as fertilizer.
Description
Technical field：
The invention belongs to storing technology field, and in particular to a kind of method of long-term storage entomopathogenic nematode.
Background technology：
Entomopathogenic nematode is biological insecticides new in the world, and with a large amount of productions are easy to, host range is wide, lethal
The fireballing advantage of insect, has been widely used in biological control (Georgis＆Manweiler, 1994 of subterranean pest-insect；
Shapiro-Ilan et al.,2002；Grewal et al.,2005；Georgis et al.,2006；Yan et al.,
2012；Shapiro-Ilan et al.,2016).Guangdong Province's living resources Applied Research Laboratory (former Guangdong Province Insects Research Institute) exists
South China golf course separates a kind of entomopathogenic nematode --- and Steinernema longicaudum X-7 strains are to a variety of Qi
Scarabaeiform, such as North China Holotrichia parallela (Holotrichia parallela Motschulsky), holotrichia oblita
The field control effects such as (Holotrichia oblita Faldermann) larva up to more than 90% (Guo et al.,
2013,2015,2016), have a extensive future.In the commercialization of entomopathogenic nematode, during the extension effective shelf of nematode
Between, it is one of key technology of nematode industrialization production in order to long-distance transportation and the Field information of Best Times.This experiment
Series of parameters research has been done in room for the shelf storage of S.longicaudum X-7 nematodes, finally gives one kind
The long-term storage method of S.longicaudum X-7 nematodes.
The content of the invention：
It is an object of the invention to provide a kind of side of the high long-term storage entomopathogenic nematode of storage period length, survival rate
Method.
The method of the long-term storage entomopathogenic nematode of the present invention, comprises the following steps：By nematode mud and fertile soil according to
Mass ratio 1:0.8~1.2 it is well mixed obtain mixture, then fill this blend into container, after being sealed with breathable sealing film
Stored；Described nematode mud is made up of entomopathogenic nematode infection period nematode and water.
Nematode density is preferably 600,000/g in described nematode mud.
Described entomopathogenic nematode is preferably Steinernema longicaudum X-7.
It is described sealed with breathable sealing film after stored, its storage temperature is preferably 5 DEG C~25 DEG C.Described storage
Temperature is more preferably 5 DEG C~15 DEG C.
Described particle diameter humous is preferably 24 mesh.
It is preferred that, the ventilated membrane that described breathable sealing film includes pp films and is arranged in the middle part of pp films.
There is the method for the present invention raw material to be easy to get, easily operation, cost is low, occupy little space, period of storage length, survival rate
High advantage.S.longicaudum X-7 nematodes press 1 with fertile soil:0.8~1:1.2 mixtures are up to after 5 DEG C of stable storages
180d, survival rate is still greater than 90%.The entomopathogenic nematode of storage, which can be directly spread fertilizer over the fields to field, to be used to prevent and treat Field Pests, together
When can also be used as fertilizer improve field fertility.
Brief description of the drawings：
Fig. 1 be entomopathogenic nematode Steinernema longicaudum X-7 respectively with diatomite, vermiculite, fertile soil
According to mass ratio 1:1 or with broken sponge according to mass ratio 3:Storage effect of 1 mixture under the conditions of 5 DEG C, 15 DEG C, 25 DEG C；Its
In, * shows significant difference (P between same sample time different disposal nematode survival rate<0.05, Tukey ' s test).
Embodiment：
Following examples are that the present invention is further illustrated, rather than limitation of the present invention.
Embodiment 1：
First, experiment material and method
The culture of 1.S.longicaudum X-7 nematodes：S.longicaudum X-7 nematode infections phases nematode (IJs,
Infective juveniles), cultivated according to the solid culture method of Korean and Japanese farmland (1996).Harvesting infection period nematode will
Nematode loaded on being placed in washing machine and being cleaned according to conventional program in gauze bag, collect washing machine drainage liquid in subsider (2.0m ×
1.0m × 0.5m) in, the collection nematode after cleaning 3 times that adds water is deposited in collection nematode mud in cotton bag.
2. nematode mud Statistics of Density：Line taking worm mud 0.1g is dissolved in 1000mL water respectively, is stirred, taken with glass bar
1mL is in tally.Every kind of nematode mud and repeated sampling 3 in each nematode suspension, in being counted under microscope, are averaged
The nematode density that value calculates in the nematode mud of harvest is 600,000/g.
3. nematode prepares formulation in proportion with different carriers：By nematode mud and diatomite (150 mesh) respectively according to mass ratio
1:0.8、1:1、1:1.2 are well mixed, and obtain nematode diatomite formulation.By nematode mud and vermiculite (200 mesh) respectively according to quality
Than 1:0.8、1:1、1:1.2 are well mixed, and obtain nematode vermiculite formulation.By nematode mud and fertile soil (24 mesh) respectively according to quality
Than 1:0.8、1:1、1:1.2 are well mixed, and obtain nematode fertile soil formulation.By nematode mud and broken sponge (d=1cm) according to quality
Than 3:1 is well mixed, and obtains the broken sponge formulation of nematode.
4. nematode formulation is dispensed：Will it is well mixed after nematode formulation according to 40g/ bottle (diatomite and vermiculite formulations),
23g/ bottles (fertile soil formulation), 13g/ bottles (broken sponge formulation) load plastic cup, and (pp materials, 200mL, upper bore is 4.5cm, bottom
The a diameter of 7cm in portion, is highly 7.5cm) in, height of all formulations in plastic cup is 5.5 ± 0.5cm.Plastic cup is used
(breathable sealing film includes pp films and the ventilated membrane that is arranged in the middle of pp films to breathable sealing film, a diameter of 1cm of ventilated membrane, thoroughly
The membrane aperture of air film is 5 μm) sealed.
5. the storage temperature of nematode formulation is set：By the processing of the formulations of various carrier different proportions be respectively placed in 5 DEG C, 15
DEG C, stored in 25 DEG C of storerooms, each processing is 30 repetitions (30 plastic cups).
6. nematode storage effect is determined：Be 0 at regular intervals, 20,40,60,80,100,120,140,160,180 days
When sampling detection nematode survival rate, 3 repetitions of each processing sampling every time, each repetition takes 3 sample counts.Sampling method
For：All nematodes in each cup will each be handled all soluble in water, 10mL is sampled after stirring in sterilizing culture dish,
Recover to count after 24h under the conditions of being placed in 25 DEG C, count the amount of survival and The dead quantity of nematode, calculate the survival rate of nematode.
7. data analysis：The percent value of nemic death rate is through arcsine arcsin (X1/2) after conversion, with SPSS softwares
(version is SPSS 16.0for Windows) carries out variance analysis, and Multiple range test examines the difference between each processing using Tukey
Different conspicuousness, level of signifiance P<0.05 (SPSS 16.0, SPSS Inc., Chicago, IL, USA).
2nd, experimental result
Nematode and three kinds of mixed ratios 1 of identical carrier:0.8、1:1、1:Between 1.2 nematode survival rate at the same temperature
Do not make significant difference (F≤4.650；Df=2,6；P≥0.06).Nematode is with each carrier according to mass ratio 1:1 and nematode and broken sea
Storage effect of the continuous mixture at 5 DEG C, 15 DEG C, 25 DEG C refers to Fig. 1.At the same temperature, the survival of each nematode diatomite formulation
Rate is substantially less than vermiculite formulation and fertile soil formulation (F >=10.368；Df=3,8；P≤0.004).S.longicaudum X-7
In 5 DEG C, 15 DEG C, 25 DEG C with vermiculite, fertile soil or broken sponge mixture to the storage of nematode without significant difference.S.longicaudum
The survival rate of X-7 nematodes nematode at 5 DEG C 180 days, 15 DEG C 80 days, 25 DEG C 20 days is still greater than 90%.
Therefore, S.longicaudum X-7 nematodes can be with vermiculite, fertile soil in mass ratio 1:0.8~1.2 mixture or with it is broken
Sponge is according to mass ratio 3:1 mixture is up to 180d after 5 DEG C of stable storages, and survival rate is still greater than 90%.
But it is due to the more difficult operation of sponge, and fertile soil can be mixed as Fertilizer application, therefore by nematode with fertile soil
Close, you can to have stored nematode well, can be killed two birds with one stone again as Fertilizer application, and fertile soil is cheap, compares vermiculite
Cheap half, therefore optimal formulation or fertile soil formulation.
Claims (7)
1. a kind of method of long-term storage entomopathogenic nematode, it is characterised in that comprise the following steps：By nematode mud and fertile soil
According to mass ratio 1:0.8~1.2 it is well mixed obtain mixture, then fill this blend into container, sealed with breathable sealing film
Stored after mouthful；Described nematode mud is made up of entomopathogenic nematode infection period nematode and water.
2. the method for long-term storage entomopathogenic nematode according to claim 1, it is characterised in that in described nematode mud
Nematode density is 600,000/g.
3. the method for long-term storage entomopathogenic nematode according to claim 1, it is characterised in that described Insect Pathogenic
Nematode is Steinernema longicaudum X-7.
4. the method for long-term storage entomopathogenic nematode according to claim 1, it is characterised in that the ventilative envelope of described use
Stored after membrana oralis sealing, its storage temperature is 5 DEG C~25 DEG C.
5. the method for long-term storage entomopathogenic nematode according to claim 4, it is characterised in that described storage temperature
For 5 DEG C~15 DEG C.
6. the method for long-term storage entomopathogenic nematode according to claim 1, it is characterised in that described is humous
Particle diameter is 24 mesh.
7. the method for long-term storage entomopathogenic nematode according to claim 1, it is characterised in that described breathable sealing
Film includes pp films and the ventilated membrane being arranged in the middle part of pp films.
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| CN201710297024.1A CN106962324A (en) | 2017-04-28 | 2017-04-28 | A kind of method of long-term storage entomopathogenic nematode |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108967360A (en) * | 2018-06-14 | 2018-12-11 | 广东省生物资源应用研究所 | A kind of method of quick collection entomopathogenic nematode |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6422801A (en) * | 1987-07-16 | 1989-01-25 | Oji Paper Co | Preservation of nematode living in insect |
| CN1335061A (en) * | 2000-07-20 | 2002-02-13 | 广东省昆虫研究所 | Storing method of entomopathogenic nematode |
| CN101978824A (en) * | 2009-06-26 | 2011-02-23 | 南开大学 | Method for preserving entomopathogenic nematode at normal temperature by using expanded vermiculite |
| CN102146343A (en) * | 2010-12-31 | 2011-08-10 | 吴海燕 | Storage method capable of keeping activity of nematode-trapping fungi for long time |
| CN106035250A (en) * | 2016-08-18 | 2016-10-26 | 浙江绿神天敌生物技术有限公司 | Entomopathogenic nematode culture process |
| CN106172259A (en) * | 2016-08-18 | 2016-12-07 | 浙江绿神天敌生物技术有限公司 | A kind of entomopathogenic nematode cleaning method |
-
2017
- 2017-04-28 CN CN201710297024.1A patent/CN106962324A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6422801A (en) * | 1987-07-16 | 1989-01-25 | Oji Paper Co | Preservation of nematode living in insect |
| CN1335061A (en) * | 2000-07-20 | 2002-02-13 | 广东省昆虫研究所 | Storing method of entomopathogenic nematode |
| CN101978824A (en) * | 2009-06-26 | 2011-02-23 | 南开大学 | Method for preserving entomopathogenic nematode at normal temperature by using expanded vermiculite |
| CN102146343A (en) * | 2010-12-31 | 2011-08-10 | 吴海燕 | Storage method capable of keeping activity of nematode-trapping fungi for long time |
| CN106035250A (en) * | 2016-08-18 | 2016-10-26 | 浙江绿神天敌生物技术有限公司 | Entomopathogenic nematode culture process |
| CN106172259A (en) * | 2016-08-18 | 2016-12-07 | 浙江绿神天敌生物技术有限公司 | A kind of entomopathogenic nematode cleaning method |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108967360A (en) * | 2018-06-14 | 2018-12-11 | 广东省生物资源应用研究所 | A kind of method of quick collection entomopathogenic nematode |
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Application publication date: 20170721 |
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