CN105706571A - Method for eliminating seed-borne grass endophytic fungi - Google Patents
Method for eliminating seed-borne grass endophytic fungi Download PDFInfo
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- CN105706571A CN105706571A CN201610126593.5A CN201610126593A CN105706571A CN 105706571 A CN105706571 A CN 105706571A CN 201610126593 A CN201610126593 A CN 201610126593A CN 105706571 A CN105706571 A CN 105706571A
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- seed
- distilled water
- grass
- ultrasonoscope
- kernel
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C1/00—Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
Abstract
The invention provides a method for eliminating seed-borne grass endophytic fungi. The method comprises the steps of 1, heating distilled water to 50+/-2 DEG C, then keeping the temperature unchanged, and placing grass seeds in the distilled water; 2, placing the distilled water which is 50+/-2 DEG C into an ultrasonoscope, and placing a container containing the seeds into the ultrasonoscope quickly for ultrasonic treatment; 3, conducting ultrasonic treatment on the infected seeds for 20-25 min, and keeping the temperature of the distilled water in the container and the ultrasonoscope at 50+/-2 DEG C during ultrasonic treatment. The method has the advantages thatendophytic fungus killing effect is good; application is easy, and seed-borne grass endophytic fungi can be killed with simple instruments under simple conditions; efficiency is high, and seed-borne grass endophytic fungi can be killed without one hour; cost is low, simple instruments and equipment are adopted, and energy consumption is low; damage to seed vigor is small, seed membrane lipid damage can not be caused easily, and seed vigor is improved to a certain extent through ultrasonic treatment.
Description
Technical field
The present invention relates to a kind of method eliminating kind and passing dogstail endogenetic fungus.
Background technology
EpichloëBelonging to endogenetic fungus can be with the dogstail-endogenetic fungus homobium of major part cold-season-type dogstail formation mutualism.Generally can produce the alkaloids secondary metabolite poisonous to domestic animal owing to endogenetic fungus infects this homobium of formation, bring immeasurable loss to the production of every country grassland agriculture.In nature, part dogstail endogenetic fungus infection rate is up to 100%, therefore research worker is in order to determine that this kind of endogenetic fungus is to the growth of host dogstail and ecological effect, it is often necessary to eliminates its existence in host, sets up the test material without endogenetic fungus and compare research.Some sterilizing methods often operate and waste time and energy and expensive at present, and part instrument cannot be popularized, and the sterilizing methods hence setting up a kind of efficient quick is imperative.Meanwhile, this endogenetic fungus carries out vertical transmission mainly by host's seed, thus starts with from propagation source and go out except endogenetic fungus, can the most effectively control its propagation and vigor.
Eliminate kind of the method passing dogstail endogenetic fungus at present and have following several: (1) hot bath sterilizing: utilize boiling water bath to add the seed-born fungi that heat extraction Interior Seed carries.Shortcoming is: degerming inefficient, and boiling water bath easily causes seed vitality to decline.(2) high temperature dry heat sterilization: be positioned in baking oven by seed, utilizes long dry heat treatment that seed is carried out sterilizing.Shortcoming is: spends the time longer, consumes energy higher, and sterilization effect is general.(3) fungicide feed treatment: infected seed is placed in certain density microbicide solution and soaks sterilizing for a long time.Shortcoming is: expensive relative to additive method, is unfavorable for the sterilization treatment of a large amount of seed.Accordingly, it would be desirable to find eliminating of a kind of excellence to plant the method passing dogstail endogenetic fungus.
Summary of the invention
In order to solve problems of the prior art, the invention provides a kind of method eliminating kind and passing dogstail endogenetic fungus, degerming efficiency reaches 95%, and will not damage seed vitality, take a short time, consumes energy low, cheap, is suitable to the sterilization treatment of a large amount of seed.
The present invention provides a kind of and eliminates kind of the method passing dogstail endogenetic fungus, and step is as follows:
(1) distilled water is heated to 50 ± 2 DEG C of insulations, Grass seed is added distilled water；
(2) in ultrasonoscope, add 50 ± 2 DEG C of distilled water, rapidly the container adding seed is put into and ultrasonoscope carries out supersound process；
(3) infected seed carries out the supersound process of 20-25 minute, and in ultrasonication, in container and ultrasonoscope, the temperature of distilled water maintains 50 ± 2 DEG C.
Heating in water bath can go out except seed-born fungi, but temperature during water-bath or time change, all can affect dogstail kind pass endogenetic fungus go out except effect, the most easily reduce seed vitality；The method of ultrasonic sterilization is not yet applied in terms of plant seed-born fungi.The method that the present invention uses low temperature water-bath to be combined with ultrasound wave processes infected seed, and strictly control sonication treatment time, the negative effect that seed vitality is caused by ultrasound wave is farthest reduced on the premise of reaching sterilizing, by dogstail endogenetic fungus bacterial bearing rate from 100% be reduced to 5% within, reached good sterilization effect.
As preferably, described dogstail endogenetic fungus is that obligatory parasitism is in gramineous grassEpichloëBelong to endogenetic fungus.
As preferably, in step (1), the temperature of distilled water is 50 DEG C.
As preferably, in step (2), the temperature of distilled water is 50 DEG C.
As preferably, in step (3), according to the difference of Grass seed mass of 1000 kernel, infected seed carrying out the supersound process of 20-25 minute, mass of 1000 kernel is light, and sonication treatment time is short, and mass of 1000 kernel weight, sonication treatment time is long.
As further preferably, when mass of 1000 kernel is less than or equal to 1.13g, sonication treatment time is 20 minutes；Mass of 1000 kernel often increases 0.15g, and sonication treatment time increases 20s accordingly, and the longest sonication treatment time must not exceed 25 minutes.
As preferably, in step (3), supersonic frequency is 40kHz.
As further preferably, after step (3), use aseptic water washing Grass seed, dry.
As preferably, described in dry for naturally drying in ventilation.
The method of the present invention has the effect that
1, effective: endogenetic fungus goes out except effective, after process, within dogstail endogenetic fungus bacterial bearing rate is reduced to 5%；
2, easy: complicated instrument and treatment conditions i.e. need not can to go out remove kind passing a dogstail fungus；
3, efficient: plus time, the sterilizing works of seed just can complete in one hour；
4, expense is relatively low: this method relates to material and is only some relatively simple instrument and equipments such as ultrasonoscope, distilled water, container, consumes energy relatively low；
5, less to seed vitality damage: the temperature owing to processing is relatively low and the time is shorter, do not easily cause the damage of seed membrane fat, and a certain degree of ultrasonic Treatment improves the vigor of seed.
Detailed description of the invention
Below example facilitates a better understanding of the present invention, but does not limit the present invention.Experimental technique in following embodiment, if no special instructions, is conventional method.Test material used in following embodiment, if no special instructions, is commercially available.
Instrument: ultrasonoscope (model: SB-5200D；Frequency: 40kHz；Power: 300W).
Kind of the method passing dogstail endogenetic fungus that eliminates of the present invention is:
(1) distilled water is heated to 50 ± 2 DEG C of insulations standby；
(2) Grass seed carried disease germs is placed in container, adds 50 ± 2 DEG C of distilled water and do not had seed, standby；
(3) in ultrasonoscope, add 50 ± 2 DEG C of distilled water, rapidly the container adding seed is put into and ultrasonoscope carries out supersound process；
(4) according to the difference (typically between 1.13g-4.45g) of Grass seed mass of 1000 kernel, infected seed carrying out the supersound process of 20-25 minute, in ultrasonication, in container and ultrasonoscope, the temperature of distilled water maintains 50 ± 2 DEG C.Mass of 1000 kernel is light, and sonication treatment time is short, and mass of 1000 kernel weight, sonication treatment time is long.The preferred process time is as follows: when mass of 1000 kernel is less than or equal to 1.13g, sonication treatment time is 20 minutes；Mass of 1000 kernel often increases 0.15g, and sonication treatment time increases 20s accordingly, and the longest sonication treatment time must not exceed 25 minutes；
(5) after supersound process completes, use aseptic water washing 3-5 time rapidly, be placed in shading ventilation and naturally dry, i.e. obtain the Grass seed without seed-born fungi.
Dogstail endogenetic fungus mentioned in the present invention refers to that obligatory parasitism is in gramineous grassEpichloëBelong to endogenetic fungus.After seed should being processed in aforementioned manners, dogstail endogenetic fungus bacterial bearing rate from 100% be reduced to 5% within, reached good sterilization effect.And not easily causing the damage of seed membrane fat, a certain degree of ultrasonic Treatment also enhances the vigor of seed, and the method for the present invention can improve Grass seed germinating energy 4-7 percentage point, improves Grass seed germination percentage 3-7 percentage point.
Embodiment 1
Elimination Herba oxytropis glabrae (Achnatherum
inebrians) method of endogenetic fungus is:
(1) distilled water is heated to 48 DEG C of insulations standby；
(2) infected seed is placed in container, adds 48 DEG C of distilled water and do not had Herba oxytropis glabrae seed, standby；
(3) in ultrasonoscope, add 48 DEG C of distilled water, rapidly the container adding Herba oxytropis glabrae seed is put into and ultrasonoscope carries out supersound process；
(4) mass of 1000 kernel of Herba oxytropis glabrae is 1.43g, according to the mass of 1000 kernel of Herba oxytropis glabrae, infected seed carries out 2 points of supersound process of 40 seconds, and in ultrasonication, in container and ultrasonoscope, the temperature of distilled water maintains 50 ± 2 DEG C.
(5) after supersound process completes, use aseptic water washing 4 times rapidly, be placed in ventilation and naturally dry, i.e. obtain the Herba oxytropis glabrae seed without seed-born fungi.
After the method for the application present invention processes Herba oxytropis glabrae seed, endogenetic fungus bacterial bearing rate is reduced to 4%, does not easily cause the damage of Herba oxytropis glabrae seed membrane fat simultaneously, and germinating energy brings up to 67-69% from 62%, and germination percentage brings up to 87%-89% from 81%.
Embodiment 2
Elimination rye grass (Lolium
perenne) method of endogenetic fungus is:
(1) distilled water is heated to 49 DEG C of insulations standby；
(2) infected seed is placed in container, adds 49 DEG C of distilled water and do not had ryegrass seed, standby；
(3) in ultrasonoscope, add 49 DEG C of distilled water, rapidly the container adding ryegrass seed is put into and ultrasonoscope carries out supersound process；
(4) mass of 1000 kernel of rye grass is 2.35g, according to the mass of 1000 kernel of rye grass, infected seed carries out the supersound process of 22 minutes 43 seconds, and in ultrasonication, in container and ultrasonoscope, the temperature of distilled water maintains 50 ± 2 DEG C.
(5) after supersound process completes, use aseptic water washing 5 times rapidly, be placed in ventilation and naturally dry, i.e. obtain the ryegrass seed without seed-born fungi.
After the method for the application present invention processes ryegrass seed, endogenetic fungus bacterial bearing rate is reduced to 5%, does not easily cause the damage of ryegrass seed film fat simultaneously, and germinating energy brings up to 73% from 68%, and germination percentage brings up to 94% from 90%.
Embodiment 3
Elimination Festuca Arundinacea (Festuca
arundinacea) method of endogenetic fungus is:
(1) distilled water is heated to 50 DEG C of insulations standby；
(2) infected seed is placed in container, adds 50 DEG C of distilled water and do not had tall fescue seed, standby；
(3) in ultrasonoscope, add 50 DEG C of distilled water, rapidly the container adding tall fescue seed is put into and ultrasonoscope carries out supersound process；
(4) mass of 1000 kernel of Festuca Arundinacea is 2.51g, according to the mass of 1000 kernel of Festuca Arundinacea, infected seed carries out the supersound process of 23 minutes 4 seconds, and in ultrasonication, in container and ultrasonoscope, the temperature of distilled water maintains 50 ± 2 DEG C.
(5) after supersound process completes, use aseptic water washing 3 times rapidly, be placed in ventilation and naturally dry, i.e. obtain the tall fescue seed without seed-born fungi.
After the method for the application present invention processes tall fescue seed, endogenetic fungus bacterial bearing rate is reduced to 4%, does not easily cause the damage of tall fescue seed film fat simultaneously, and germinating energy brings up to 69% from 65%, and germination percentage brings up to 91% from 88%.
Embodiment 4
Elimination lyme grass (Elymus
dahuricus) method of endogenetic fungus is:
(1) distilled water is heated to 51 DEG C of insulations standby；
(2) infected seed is placed in container, adds 51 DEG C of distilled water and do not had lyme grass seed, standby；
(3) in ultrasonoscope, add 51 DEG C of distilled water, rapidly the container adding lyme grass seed is put into and ultrasonoscope carries out supersound process；
(4) mass of 1000 kernel of lyme grass is 4.21g, according to the mass of 1000 kernel of lyme grass, infected seed carries out the supersound process of 25 minutes, and in ultrasonication, in container and ultrasonoscope, the temperature of distilled water maintains 50 ± 2 DEG C.
(5) after supersound process completes, use aseptic water washing 5 times rapidly, be placed in ventilation and naturally dry, i.e. obtain the lyme grass seed without seed-born fungi.
After the method for the application present invention processes lyme grass seed, endogenetic fungus bacterial bearing rate is reduced to 4%, does not easily cause the damage of lyme grass seed membrane fat simultaneously, and germinating energy brings up to 58% from 54%, and germination percentage brings up to 85% from 82%.
Embodiment 5
Elimination Festuca sinensis (Festuca
sinensis) method of endogenetic fungus is:
(1) distilled water is heated to 52 DEG C of insulations standby；
(2) infected seed is placed in container, adds 52 DEG C of distilled water and do not had Festuca sinensis seed, standby；
(3) in ultrasonoscope, add 52 DEG C of distilled water, rapidly the container adding Festuca sinensis seed is put into and ultrasonoscope carries out supersound process；
(4) mass of 1000 kernel of Festuca sinensis is 0.87g, according to the mass of 1000 kernel of Festuca sinensis, infected seed carries out the supersound process of 20 minutes, and in ultrasonication, in container and ultrasonoscope, the temperature of distilled water maintains 50 ± 2 DEG C.
(5) after supersound process completes, use aseptic water washing 3 times rapidly, be placed in ventilation and naturally dry, i.e. obtain the Festuca sinensis seed without seed-born fungi.
After the method for the application present invention processes Festuca sinensis seed, endogenetic fungus bacterial bearing rate is reduced to 3%, does not easily cause the damage of Festuca sinensis seed membrane fat simultaneously, and germinating energy brings up to 45% from 41%, and germination percentage brings up to 81% from 76%.
Embodiment 6
Elimination wild barley (Hordeum
brevisubulatum) method of endogenetic fungus is:
(1) distilled water is heated to 50 DEG C of insulations standby；
(2) infected seed is placed in container, adds 50 DEG C of distilled water and do not had wild barley seed, standby；
(3) in ultrasonoscope, add 50 DEG C of distilled water, rapidly the container adding wild barley seed is put into and ultrasonoscope carries out supersound process；
(4) mass of 1000 kernel of wild barley is 1.54g, according to the mass of 1000 kernel of wild barley, infected seed carries out the supersound process of 20 minutes 55 seconds minutes, and in ultrasonication, in container and ultrasonoscope, the temperature of distilled water maintains 50 ± 2 DEG C.
(5) after supersound process completes, use aseptic water washing 4 times rapidly, be placed in ventilation and naturally dry, i.e. obtain the wild barley seed without seed-born fungi.
After the method for the application present invention processes wild barley seed, endogenetic fungus bacterial bearing rate is reduced to 4%, does not easily cause the damage of wild barley seed membrane fat simultaneously, and germinating energy brings up to 58% from 53%, and germination percentage brings up to 87% from 81%.
Embodiment 7
Elimination plumage thatch (Achnatherum
sibiricum) method of endogenetic fungus is:
(1) distilled water is heated to 50 DEG C of insulations standby；
(2) infected seed is placed in container, adds 50 DEG C of distilled water and do not had plumage thatch seed, standby；
(3) in ultrasonoscope, add 50 DEG C of distilled water, rapidly the container adding plumage thatch seed is put into and ultrasonoscope carries out supersound process；
(4) mass of 1000 kernel of plumage thatch is 2.78g, according to the mass of 1000 kernel of plumage thatch, infected seed carries out the supersound process of 23 minutes 40 seconds, and in ultrasonication, in container and ultrasonoscope, the temperature of distilled water maintains 50 ± 2 DEG C.
(5) after supersound process completes, use aseptic water washing 4 times rapidly, be placed in ventilation and naturally dry, i.e. obtain the plumage thatch seed without seed-born fungi.
After the method for the application present invention processes plumage thatch seed, endogenetic fungus bacterial bearing rate is reduced to 4%, does not easily cause the damage of plumage thatch seed membrane fat simultaneously, and germinating energy brings up to 43% from 38%, and germination percentage brings up to 79% from 74%.
Last it is noted that the foregoing is only the preferred embodiments of the present invention, it is not limited to the present invention, although the present invention being described in detail with reference to previous embodiment, for a person skilled in the art, technical scheme described in foregoing embodiments still can be modified by it, or wherein portion of techniques feature is carried out equivalent.All within the spirit and principles in the present invention, any modification, equivalent substitution and improvement etc. made, should be included within the scope of the present invention.
Claims (9)
1. one kind eliminates kind of the method passing dogstail endogenetic fungus, it is characterised in that: step is as follows:
(1) distilled water is heated to 50 ± 2 DEG C of insulations, Grass seed is added distilled water；
(2) in ultrasonoscope, add 50 ± 2 DEG C of distilled water, rapidly the container adding seed is put into and ultrasonoscope carries out supersound process；
(3) infected seed carries out the supersound process of 20-25 minute, and in ultrasonication, in container and ultrasonoscope, the temperature of distilled water maintains 50 ± 2 DEG C.
Method the most according to claim 1, it is characterised in that: described dogstail endogenetic fungus is that obligatory parasitism is in gramineous grassEpichloëBelong to endogenetic fungus.
Method the most according to claim 1, it is characterised in that: in step (1), the temperature of distilled water is 50 DEG C.
Method the most according to claim 1, it is characterised in that: in step (2), the temperature of distilled water is 50 DEG C.
Method the most according to claim 1, it is characterised in that: in step (3), according to the difference of Grass seed mass of 1000 kernel, infected seed carrying out the supersound process of 20-25 minute, mass of 1000 kernel is light, and sonication treatment time is short, and mass of 1000 kernel weight, sonication treatment time is long.
Method the most according to claim 5, it is characterised in that: when mass of 1000 kernel is less than or equal to 1.13g, sonication treatment time is 20 minutes；Mass of 1000 kernel often increases 0.15g, and sonication treatment time increases 20s accordingly, and the longest sonication treatment time must not exceed 25 minutes.
Method the most according to claim 1, it is characterised in that: in step (3), supersonic frequency is 40kHz.
8. according to the arbitrary described method of claim 1-7, it is characterised in that: after step (3), use aseptic water washing Grass seed, dry.
Method the most according to claim 8, it is characterised in that dry described in: for naturally drying in ventilation.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201610126593.5A CN105706571A (en) | 2016-03-07 | 2016-03-07 | Method for eliminating seed-borne grass endophytic fungi |
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| CN201610126593.5A CN105706571A (en) | 2016-03-07 | 2016-03-07 | Method for eliminating seed-borne grass endophytic fungi |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106305782A (en) * | 2016-08-23 | 2017-01-11 | 兰州大学 | Method for preventing and removing Achnatherum inebrians-endophytic fungi symbiont by virtue of chemical agent |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102326514A (en) * | 2011-11-01 | 2012-01-25 | 四川中测科技投资有限公司 | Tea tree explant mercury-free disinfectant and explant disinfection method |
| CN102498793A (en) * | 2011-10-22 | 2012-06-20 | 中国科学院合肥物质科学研究院 | Method for removing endophytes from pistacia chinensis bunge seeds and quickly germinating pistacia chinensis bunge seeds |
-
2016
- 2016-03-07 CN CN201610126593.5A patent/CN105706571A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102498793A (en) * | 2011-10-22 | 2012-06-20 | 中国科学院合肥物质科学研究院 | Method for removing endophytes from pistacia chinensis bunge seeds and quickly germinating pistacia chinensis bunge seeds |
| CN102326514A (en) * | 2011-11-01 | 2012-01-25 | 四川中测科技投资有限公司 | Tea tree explant mercury-free disinfectant and explant disinfection method |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106305782A (en) * | 2016-08-23 | 2017-01-11 | 兰州大学 | Method for preventing and removing Achnatherum inebrians-endophytic fungi symbiont by virtue of chemical agent |
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Application publication date: 20160629 |
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