CN105973632A - Plant root secreta generation and collection device and collection method thereof - Google Patents
Plant root secreta generation and collection device and collection method thereof Download PDFInfo
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- CN105973632A CN105973632A CN201610292586.2A CN201610292586A CN105973632A CN 105973632 A CN105973632 A CN 105973632A CN 201610292586 A CN201610292586 A CN 201610292586A CN 105973632 A CN105973632 A CN 105973632A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/02—Devices for withdrawing samples
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- Chemical & Material Sciences (AREA)
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Abstract
The invention provides a plant root secreta generation and collection device and a collection method thereof. The device comprises a plant growth device, a root growth guiding device and a root elution and secreta collection device. The root growth guiding device comprises a net structure and a plastic separator plate. The collection method is a plant root secreta collection method under soil stress under natural conditions and does not interfere root growth and damage a rhizosphere environment. The collection method comprises root secreta effective expression without outside interference after single pathogen infection and monitoring of root secreta change after rhizosphere pathogen infection.
Description
Technical field
The present invention relates to technical field of plant protection, particularly relate to a kind of secretions from plant roots and occur and receive
Acquisition means and collection method thereof.
Background technology
In plant growth and development process, root system is as the contact portion of plant Yu soil, not only from soil
Middle absorption moisture and nutrient, and by the way of root is secreted, around root, discharge some inorganic ionss and have
Machine compound, these materials are referred to as root exudates.Having numerous studies to show, root exudates affects
To soil biology environmental change and allelopathy, Different Crop is on soil microbial community impact difference.
Crop is played an important role in soil microbial community by root exudates, and Different Crop root system divides
The component content of secretion is different, and same origin is different by the component content of root exudates after pathogen infection,
Thus change the growth metabolism of soil microbial community.
At present, collect secretions from plant roots and typically use solution collecting method, and by plant growing in nutrition
In liquid or Nutrition Soil, proceeding to process in solution, after processing a period of time, collection processes solution and is root system
Secretions.This method has the drawback that: the plant being grown in nutritional solution or Nutrition Soil lacks after taking out
Few natural conditions select, and coerce without soil and are difficult to the practical situation of root system secretion under reaction edaphic condition, this
It is because that soil constituent is complicated, the variation of nutrient existence form, such as Chinese patent CN102636373A,
CN204630769U, 20052023038.7；Chinese patent 200610123460.9 discloses a kind of earth culture
Secretions from plant roots collection method, its shortcoming is the polytetrafluoroethylene film that 1. root systems are selected by place
Quality is hard, and root system is more difficult to be passed through, and 2. washs with too much solution, some organic acid in solution, inorganic from
The active ion species such as son, cation, anion easily react with root exudates matter, impact test knot
Really, 3. the mineral in soil and microorganism composition flow in collecting box with water, if 4. carrying out pathogen infection
Test, experimental condition is difficult to sterile working, it is impossible to ensure pathogen purity.With Chinese patent
201110004588.4 compare, and this patent main material selects politef, energy high temperature high voltage resistant, uses
Sterilizing before cultivating；Before plant moves into growth device for plant, agar will be filled in root growth guide
Or Nutrient medium is for fungus or bacterial reproduction, in order to root exudates after pathogenic bacterial infection plant
Collect.
Therefore, one is found under field conditions (factors), the collection of secretions from plant roots in soil stress procedure
Method is critically important.After without pathogen infection process single under the conditions of external interference, root exudates
Effective expression.Work out not disturbance root growth, do not destroy rhizospheric environment, monitor rhizosphere pathogen infection
The change of rear root exudates, the method and device effectively collecting root exudates is significant.Mesh
Before, collection method and device about this root exudates have not been reported both at home and abroad.
Summary of the invention
The technical problem to be solved is to provide one not disturbance root growth, does not destroy rhizosphere ring
The device and method of root exudates change behind border, at any time monitoring rhizosphere pathogen infection, it is therefore intended that gram
Take that root system in existing method is perishable, easily polluted by microorganism, sampling easily injures root system, can not take continuously
The drawbacks such as sample, it is provided that one under field conditions (factors), can the root system of plant of continuous sampling in soil stress procedure
Secretions collection method.After without pathogen infection under the conditions of external interference, root exudates effective
Express.
For solving above-mentioned technical problem, the invention provides a kind of secretions from plant roots and occur and collect dress
Putting, it includes growth device for plant, root growth guide and root system eluting and secretions collection device,
Described growth device for plant loads Nutrition Soil for plant field planting to be measured and nutrition supply.
Wherein, described root growth guide is made up of network structure and plastic septum, described netted knot
Structure pore size isPlastic septum is and the plastic septum of described growth device for plant bottom mating.
Wherein, after described growth device for plant is connected with root growth guide, collect plant root exudation
During thing, network structure is 0.5cm thickness agar；Collect its root system secretion after fungus or bacterial invasion plant
During thing, for connecing bacterium 0.5cm thickness culture medium (Herba bromi japonici, PDA, or NA etc.) in network structure.
Wherein, described Nutrition Soil need to carry out autoclave sterilization, and actual conditions is 120 DEG C of sterilizing 20min.
Wherein, described secretions from plant roots occurs and collection device is also required to high temperature sterilize actual conditions and is
120 DEG C of sterilizing 20min.
Wherein, described root system eluting and secretions collection device are furnished with two models of 500mL and 200mL
Catcher, collects plant such as Semen Maydis or the bigger plant 500mL catcher of well developed root system, collects seedling stage
Crop 200mL, selects different catcher according to experimental conditions.Root system eluting and secretions collection device
It is furnished with water inlet and outlet, it is simple to the collection of root exudates.
Present invention also offers a kind of employing above-mentioned secretions from plant roots generation and collection device carries out earth culture
The method that secretions from plant roots is collected, plant grows in growth device for plant, owing to growing space limits
And action of gravity, root system of plant passes root growth guide, when root system guides dress through root growth
Postpone, chain root system eluting and secretions collection device, after collection device accumulates a certain amount of root system
Can be carried out root exudates to collect.
Wherein, the method that described secretion from root system of plant in pot is collected further particularly as follows:
The first step, loads root growth guide in growth device for plant, after high temperature sterilize, ultra-clean
Pour the solid medium of thawing on workbench into, after solidification to be cooled, access plant to be infected fungus or
Bacteria pathogeny bacterium, upper end sealed membrane seals, and is placed in incubator cultivation 7-10d, treats that strain covers with whole
Take out standby after individual culture medium；
Second step, pours into bactericidal nurishing soil in the growth device for plant that step 1 obtains, cultivates in advance
Root exudates plant to be collected is colonizated in Middle nutrition soil, determines field planting number according to plant size；
3rd step, after after 5-10d, observation of plant root system arrives network structure by solid medium, takes off
Plastic septum, chains root system eluting and secretions collection device, root system eluting and secretions collection device
In fill quartz sand etc.；
4th step, until root system of plant gather in root system eluting and secretions collection device a certain amount of after, will
Quartz is removed, and loads sterilized water, collect root exudates after collection device sterilizing.
Beneficial effects of the present invention:
The present invention provide not disturbance root growth, do not destroy rhizospheric environment, at any time monitoring rhizosphere pathogen
Infect rear root exudates change device and method, it is therefore intended that overcome root system in existing method perishable,
Easily polluted by microorganism, sampling easily injures root system, can not the drawback such as continuous sampling, it is provided that a kind of at nature
Under the conditions of, can the secretions from plant roots collection method of continuous sampling in soil stress procedure.Including without outward
Under boundary's disturbed condition after pathogen infection, the effective expression of root exudates.
Accompanying drawing explanation
Fig. 1 is that the root exudates of the present invention occurs and collection device figure.
1-growth device for plant；2-root system eluting and secretions collection device；3-network structure；4-plastic partition
Plate.
Detailed description of the invention
The invention provides a kind of secretions from plant roots to occur and collection device, it includes growth device for plant,
Root growth guide and root system eluting and secretions collection device, described growth device for plant loads nutrition
Soil is for plant field planting to be measured and nutrition supply.
It is positioned at above root growth guide vegetation period of described, below root growth guide
Secretions collection device is installed.
Described root growth guide is made up of network structure and plastic septum, described network structure aperture
Size isPlastic septum is and the plastic septum of described growth device for plant bottom mating.
After described growth device for plant is connected with root growth guide, when collecting secretions from plant roots,
It is 0.5cm thickness agar in network structure；After collection fungus or bacterial invasion plant during its root exudates,
For connecing bacterium 0.5cm thickness culture medium (Herba bromi japonici, PDA, or NA etc.) in network structure.
Described Nutrition Soil need to carry out autoclave sterilization, and actual conditions is 120 DEG C of sterilizing 20min.
It is 120 DEG C that the generation of described secretions from plant roots and collection device are also required to high temperature sterilize actual conditions
Sterilizing 20min.
Described root system eluting and secretions collection device are furnished with two catchers of 500mL and 200mL, receive
The plant such as Semen Maydis of collection well developed root system or bigger plant 500mL catcher, collect crop in seedling stage and use
200mL, according to experimental conditions selection device.Root system eluting and secretions collection device are furnished with water inlet
And outlet, it is simple to the collection of root exudates.
Present invention also offers a kind of employing above-mentioned secretions from plant roots generation and collection device carries out earth culture
The method that secretions from plant roots is collected, plant grows in growth device for plant, owing to growing space limits
And action of gravity, root system of plant passes root growth guide, when root system guides dress through root growth
Postpone, chain root system eluting and secretions collection device, after collection device accumulates a certain amount of root system
Can be carried out root exudates to collect.
The method that described secretion from root system of plant in pot is collected further particularly as follows:
The first step, loads root growth guide in growth device for plant, after high temperature sterilize, ultra-clean
Pour the solid medium of thawing on workbench into, after solidification to be cooled, access plant to be infected fungus or
Bacteria pathogeny bacterium, upper end sealed membrane seals, and is placed in incubator cultivation 7-10d, treats that strain covers with whole
Take out standby after individual culture medium；
Second step, pours into bactericidal nurishing soil in the growth device for plant that step 1 obtains, cultivates in advance
Root exudates plant to be collected is colonizated in Middle nutrition soil, determines field planting number according to plant size；
3rd step, after after 5-10d, observation of plant root system arrives network structure by solid medium, takes off
Plastic septum, chains root system eluting and secretions collection device, root system eluting and secretions collection device
In fill quartz sand etc.；
4th step, until root system of plant gather in root system eluting and secretions collection device a certain amount of after, will
Quartz is removed, and loads sterilized water, collect root exudates after collection device sterilizing.
Embodiment and accompanying drawing is below used to describe embodiments of the present invention in detail, whereby to the present invention such as
What application technology means solves technical problem, and the process that realizes reaching technique effect can fully understand also
Implement according to this.
Case 1: gold root exudates little to black shank susceptible variety is collected
Root growth guide is loaded in growth device for plant 1, high temperature sterilize (120 DEG C, 20min)
After, superclean bench is poured into the agar solution of thawing, standby after cooled and solidified, by bactericidal nurishing soil
Pouring in growth device for plant 1, the little gold cultivated in advance is colonizated in Nutrition Soil, according to cigarette strain size
Determine field planting number.Whole device is put into phjytotron cultivate, under gravity, tobacco root
Growth downwards, after 20d, root system arrives the network structure 3 of root growth guide by solid medium,
Take off plastic septum 4, chain root system eluting and secretions collection device 2.At root system eluting and secretions
Collection device is filled quartz sand etc., with aluminium-foil paper parcel root system eluting and secretions collection device.Treat little
Gold root system gather in root system eluting and secretions collection device a certain amount of after, by quartz remove, receive
Load sterilized water after acquisition means sterilizing, collect root exudates.Water outlet is opened when collecting root exudates
Mouthful, little gold root exudates can be collected, by the root exudates concentrated by rotary evaporation collected or freezing
It is dried, after crossing 0.22 μm filter membrane, i.e. can be used for bioassay.To continue to collect root exudates,
Directly add sterilized water at water inlet.
Case 2: the Nicotiana tabacum L. little gold root exudates infecting tobacco black shank bacterium is collected
Root growth guide is loaded in growth device for plant 1, high temperature sterilize (120 DEG C, 20min)
After, superclean bench is poured into the oat medium of thawing, cooled and solidified is followed by tobacco black shank bacterium,
Sealed membrane sealing is placed in 28 DEG C of incubators cultivation, treats that black shank bacterium covers with whole culture medium after 7-10d
Rear taking-up, standby.Being poured in growth device for plant 1 by bactericidal nurishing soil, the little gold cultivated in advance is fixed
Plant in Nutrition Soil, determine field planting number according to cigarette strain size.Whole device is put into phjytotron training
Supporting, under gravity, tobacco root grows downwards, and after 20d, root system arrives root by solid medium
The network structure 3 of system's growth guide, takes off plastic septum 4, chains root system eluting and secretions is received
Acquisition means.In root system eluting and secretions collection device 2, fill quartz sand etc., wrap up root with aluminium-foil paper
It is eluting and secretions collection device.Treat that little gold root system is long-pending in root system eluting and secretions collection device
Poly-a certain amount of after, quartz is removed, loads sterilized water after collection device sterilizing, collect root exudates.
Open outlet when collecting root exudates, little gold root exudates can be collected, will collect
Root exudates concentrated by rotary evaporation or lyophilization, cross after 0.22 μm filter membrane, i.e. can be used for bioassay.
To continue to collect root exudates, directly add sterilized water at water inlet.
All above-mentioned primary implement these intellectual properties, do not set limit other forms enforcement this
Plant new product and/or new method.Those skilled in the art will utilize this important information, and foregoing is revised,
To realize similar implementation status.But, all modifications or transformation belong to reservation based on new product of the present invention
Right.
The above, be only presently preferred embodiments of the present invention, is not that the present invention makees other form
Restriction, any those skilled in the art are changed possibly also with the technology contents of the disclosure above
Or it is modified as the Equivalent embodiments of equivalent variations.But every without departing from technical solution of the present invention content, depend on
Any simple modification, equivalent variations and remodeling above example made according to the technical spirit of the present invention,
Still fall within the protection domain of technical solution of the present invention.
Claims (8)
1. a secretions from plant roots occurs and collection device, it is characterised in that: include growth device for plant,
Root growth guide and root system eluting and secretions collection device, described growth device for plant loads nutrition
Soil is for plant field planting to be measured and nutrition supply.
2. secretions from plant roots as claimed in claim 1 occurs and collection device, it is characterised in that:
Described root growth guide is made up of network structure and plastic septum, described network structure pore size
ForPlastic septum is and the plastic septum of described growth device for plant bottom mating.
3. secretions from plant roots as claimed in claim 1 or 2 occurs and collection device, and its feature exists
In: after described growth device for plant is connected with root growth guide, when collecting secretions from plant roots,
It is 0.5cm thickness agar in network structure；After collection fungus or bacterial invasion plant during its root exudates,
For connecing bacterium 0.5cm thickness culture medium in network structure.
4. the secretions from plant roots as described in claims 1 to 3 occurs and collection device, and its feature exists
In: described Nutrition Soil need to carry out autoclave sterilization, and actual conditions is 120 DEG C of sterilizing 20min.
5. the secretions from plant roots as described in Claims 1-4 occurs and collection device, and its feature exists
In: it is 120 DEG C that the generation of described secretions from plant roots and collection device are also required to high temperature sterilize actual conditions
Sterilizing 20min.
6. the secretions from plant roots as described in claim 1 to 5 occurs and collection device, and its feature exists
In: described root system eluting and secretions collection device are furnished with two catchers of 500mL and 200mL, receive
The plant such as Semen Maydis of collection well developed root system or bigger plant 500mL catcher, collect crop in seedling stage and use
200mL, according to experimental conditions selection device.Root system eluting and secretions collection device are furnished with water inlet
And outlet, it is simple to the collection of root exudates.
7. one kind uses the generation of above-mentioned secretions from plant roots and collection device to carry out earth culture plant root exudation
The method that thing is collected, it is characterised in that: plant grows in growth device for plant, owing to growing space limits
And action of gravity, root system of plant passes root growth guide, when root system guides dress through root growth
Postpone, chain root system eluting and secretions collection device, after collection device accumulates a certain amount of root system
Can be carried out root exudates to collect.
8. the method that secretion from root system of plant in pot is collected as claimed in claim 7, it is characterised in that bag
Include:
The first step, loads root growth guide in growth device for plant, after high temperature sterilize, ultra-clean
Pour the solid medium of thawing on workbench into, after solidification to be cooled, access plant to be infected fungus or
Bacteria pathogeny bacterium, upper end sealed membrane seals, and is placed in incubator cultivation 7-10d, treats that strain covers with whole
Take out standby after individual culture medium；
Second step, pours into bactericidal nurishing soil in the growth device for plant that step 1 obtains, cultivates in advance
Root exudates plant to be collected is colonizated in Middle nutrition soil, determines field planting number according to plant size；
3rd step, after after 5-10d, observation of plant root system arrives network structure by solid medium, takes off
Plastic septum, chains root system eluting and secretions collection device, root system eluting and secretions collection device
In fill quartz sand etc.；
4th step, until root system of plant gather in root system eluting and secretions collection device a certain amount of after, will
Quartz is removed, and loads sterilized water, collect root exudates after collection device sterilizing.
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| CN201610292586.2A CN105973632B (en) | 2016-05-05 | 2016-05-05 | Secretions from plant roots generation and collection device and its collection method |
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|---|---|---|---|
| CN201610292586.2A CN105973632B (en) | 2016-05-05 | 2016-05-05 | Secretions from plant roots generation and collection device and its collection method |
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| CN105973632A true CN105973632A (en) | 2016-09-28 |
| CN105973632B CN105973632B (en) | 2019-01-25 |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107976330A (en) * | 2017-11-21 | 2018-05-01 | 东北师范大学 | The collection device in situ and method of secretion from root system of plant in pot and rhizosphere soil |
| CN109221112A (en) * | 2018-11-12 | 2019-01-18 | 哈尔滨工业大学 | A kind of allelochemical plant growth regulator and its method for extraction and purification |
| CN110531028A (en) * | 2019-04-22 | 2019-12-03 | 贵阳学院 | Polygonum capitatum root exudates extracting method and device |
| CN110672351A (en) * | 2019-11-08 | 2020-01-10 | 华南农业大学 | Device and method for collecting root exudates of gramineous plants |
| CN113049300A (en) * | 2021-02-09 | 2021-06-29 | 湖南农业大学 | Separation and collection device and method for plant root exudates |
| CN114375695A (en) * | 2021-12-28 | 2022-04-22 | 四川农业大学 | In-situ collecting device and method for hygrophyte root exudates |
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| CN114375695A (en) * | 2021-12-28 | 2022-04-22 | 四川农业大学 | In-situ collecting device and method for hygrophyte root exudates |
| CN114375695B (en) * | 2021-12-28 | 2022-10-21 | 四川农业大学 | In-situ collecting device and method for hygrophyte root exudates |
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| Publication number | Publication date |
|---|---|
| CN105973632B (en) | 2019-01-25 |
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