CN103776662B - A kind of xerophyte root exudates gathering-device and collection method thereof - Google Patents
A kind of xerophyte root exudates gathering-device and collection method thereof Download PDFInfo
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- CN103776662B CN103776662B CN201410029083.7A CN201410029083A CN103776662B CN 103776662 B CN103776662 B CN 103776662B CN 201410029083 A CN201410029083 A CN 201410029083A CN 103776662 B CN103776662 B CN 103776662B
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Abstract
The invention provides a kind of xerophyte root exudates gathering-device, comprise a cultivation bucket, described cultivation bucket, cultivate bucket bottom surface and be communicated with a rubber tube, the described rubber tube other end is connected with syringe, soil vermiculite Mixed culture matrix layer and filtering layer is equipped with from top to bottom successively in described cultivation bucket, in described cultivation bucket, cloth is also provided with irrigation pipe, the present invention devises a kind of xerophyte root exudates collection method according to above-mentioned gathering-device, the present invention is applicable to plant under soil cultivation condition, especially the research of xerophyte root exudates, to cultivate with simple matrix with water planting and compare, more can reflect the actual state of secretions from plant roots under natural conditions, adopt this harvester under the prerequisite ensureing plant normal growth simultaneously, not disturbance root growth, do not destroy rhizospheric environment, avoid the root system damage because causing when traditional earth culture method collects root exudates and bleeding interference.
Description
Technical field
The present invention relates to a kind of xerophyte root exudates gathering-device and collection method thereof.
Background technology
At present, the collection of root exudates mainly contains water culture, substrate culture collecting method, earth culture method.Water culture does not need special gathering-device, but the general roots development of hydroponic plant is less, does not have mechanical resistance, has larger difference with the ventilatory conditions grown under real earth culture condition, Nutrient distribution, root system branched structure.Therefore water culture cannot obtain xerophytic root exudates.Matrix cultivation can ensure the ventilation of root system, also has certain mechanical resistance, but there is larger difference with the root exudates under state of nature.
Tradition earth culture method is collected root exudates and is mainly adopted two kinds of methods, and one is after earth culture plant growth a period of time, is taken out by its root system, and with distilling the lower rhizosphere soil of washing, centrifugal or filtration after concussion, gained is root exudates.Two is taken out from soil by the plant growing into certain phase, and rinsed well by its root system surface soil layer, then use distilled water drip washing root system, leacheate is as root exudates.In whole operating process, root system very easily sustains damage, and can comprise inclusions and the bleeding sap of a lot of root system itself in the solution collected, and this method be generally only applicable to small plant or seedling stage plant.Except above several method, according to the difference of research purpose and research object, the special device of some researcher's designed, designeds collects root exudates.As continuous root exudates collection system (TangandYoung, 1982), this device utilizes macroporous adsorbing resin for purification root exudates, achieves organic enrichment and is separated, but the more difficult control of the flow of nutrient solution.Recently, Oburger etc. (2013) devise the root case of a set of novelty to collect the root exudates of earth culture maize seedling.This device neither can destroy the self-sow situation between root system and soil, also can selection site preferably, but cannot collect the root exudates of block root or tuberous plant.
In sum, root exudates gathering-device used at present is all not suitable for the collection of xerophyte root exudates, which greatly limits the research to xerophyte root exudates.The present invention proposes a kind of gathering-device and method of xerophyte root exudates, be expected to the powerful becoming the research of xerophyte root exudates.
Summary of the invention
The present invention improves the problems referred to above, and namely the technical problem to be solved in the present invention is the collection that existing root exudates gathering-device is all not suitable for xerophyte root exudates, limits the research to xerophyte root exudates.
First specific embodiments of the present invention is: a kind of xerophyte root exudates gathering-device, comprise a cultivation bucket, described cultivation bucket bottom is funnel-form, described cultivation bucket bottom surface is communicated with a rubber tube, the described rubber tube other end is connected with syringe, soil vermiculite Mixed culture matrix layer and filtering layer is equipped with from top to bottom successively in described cultivation bucket, the irrigation pipe inserting soil vermiculite Mixed culture matrix layer is laid with in described cultivation bucket, Mixed culture matrix layer upper surface is exposed in described irrigation pipe upper end, described irrigation pipe is pvc pipe, pvc pipe lower end absorbent cotton seals, the one-sided boring of pipe shaft is also enclosed with 2 layers of gauze, matrix 3 ~ 4cm is exposed in irrigation pipe upper end, described filtering layer comprises sandy layer and is positioned at the nonwoven layer below sandy layer.
Further, in the middle part of described rubber tube, there is liquid stopping clamp.
Second specific embodiments of the present invention is: a kind of xerophyte root exudates collection method, utilizes a kind of xerophyte root exudates gathering-device as claimed in claim 1, comprises the following steps:
(1) first make root exudates gathering-device, in cultivation bucket, fill filtering layer and soil vermiculite Mixed culture matrix;
(2) irrigation pipe is buried, and planting plants;
(3) in plant incubation, the regular irrigation soils leaching liquor of irrigation pipe is passed through, to guarantee the normal growth of plant;
(4) before collecting root exudates, the leaching liquor slowly pouring into source identical with culture matrix soil from irrigation pipe, to there are flowing out in rubber tube, drains the solution of outflow gently with syringe, extracts solution out and is root exudates;
(5) root exudates step (4) collected is the filtering with microporous membrane of 0.22 μm through diameter, and preserves stand-by under the root exudates after filtration is transferred to 4 DEG C of conditions.
Further, described leaching liquor presses the water and soil mass ratio of 2:1 by the soil in source identical with culture matrix, shakes 4 hours and obtains after overnight stand.
Further, described culture matrix is the potpourri of soil and vermiculite, volume ratio is 1:0.5 ~ 1:1.5, filtering layer by clean oven dry nonwoven fabrics and lay and nonwoven fabrics on sand form, sand is 0.5 ~ 3kg, particle diameter 0.5 ~ 1mm, described nonwoven fabrics filling thickness is 3 ~ 8cm, limpid to ensure that root exudates collects liquid.
Further, described cultivation bucket is organic glass bucket, and organic glass bucket inwall is frosted structure, bottomless, high 20 ~ 50cm, diameter 15 ~ 30cm; Bucket shoulder and bucket stem are upside down funnel shape, bucket shoulder height 3 ~ 8cm, bucket stem 3 ~ 8cm, diameter 2 ~ 4cm, and bucket stem end closes up and is convenient to the installation of rubber tube.
Compared with prior art, the present invention has following beneficial effect:
(1) the present invention is applicable to the research of plant (especially xerophyte) root exudates under soil cultivation condition.To cultivate with simple matrix with water planting and compare, more can reflect the actual state of secretions from plant roots under natural conditions.
(2) the present invention adopts the potpourri of soil and vermiculite as culture matrix, both ensure that the nutritional need of plant normal growth, and has oozed under being conducive to again moisture, can ensure effective collection of root exudates, close to state of nature.Difference needs the collection of water type secretions from plant roots to realize by the ratio of adjustment soil and vermiculite.
(3) the present invention is under the prerequisite ensureing plant normal growth, not disturbance root growth, does not destroy rhizospheric environment, avoids the root system damage because causing when traditional earth culture method collects root exudates and bleeding interference.
(4) the present invention utilizes and irrigates through the pvc pipe of special processing, and guarantee moisture slowly to plant root infiltration from pipe, this makes the collection of root exudates more efficient.Secondly the use of pvc pipe ensure that the gas penetration potential of plant root.
(5) the present invention adopts clean fine sand and nonwoven fabrics as filtration unit, avoids the loss of soil and vermiculite, is conducive to the collection of percolate.
(6) the present invention utilizes syringe to carry out the collection of root exudates, can need to extract root exudates accurately according to experiment, compared with the peristaltic pump in conventional apparatus, extracting dynamics more easy to control, can not spoiled soil structure, the fluid sample simultaneously obtained directly can be crossed film process and analyzes for HPLC.
(7) the present invention's solution of filling with when root exudates is collected is soil extraction instead of tap water, this avoid because root exudates collects the soil nutrient loss caused, the impact of root exudates collection procedure on plant growth is dropped to minimum.
Accompanying drawing explanation
Fig. 1 is xerophyte root exudates gathering-device structural representation of the present invention.
Embodiment
Below in conjunction with the drawings and specific embodiments, the present invention will be further described in detail.
As shown in Figure 1, culture apparatus is the organic glass bucket 2 of inwall frosted, and fill the potpourri 3 that culture matrix is soil and vermiculite, research object is for treating measuring plants 1.Device for irrigating is the pvc pipe 4 through special processing, and fill in organic glass bucket and eluriate clean sand 5, nonwoven fabrics 6 is filled at bucket stem place.Control the collection of root exudates with agrafe 7, rubber tube 8 other end is connected with gathering-device and extracting syringe 9.
The collection of glutinous rehmannia root exudates:
First customize high 40 ㎝, thick 0.2 ㎝, ladle body 28 ㎝, diameter 21 centimetres, 4 ㎝ takeed on by bucket, and the bottomless frosted interior organic glass bucket of bucket stem 8 diameter 4 ㎝, nonwoven fabrics and sand are filled, nonwoven fabrics is filled in barrel stem, thickness 5 ㎝.Sand covers on nonwoven fabrics, sand 2 ㎏, particle diameter 1 ㎜.Then load culture matrix, soil vermiculite volume ratio is 1:1, fills up culture vessel, and soil takes from glutinous rehmannia planting base, Wen County, Henan.Be that square symmetry buries pvc pipe during filling, pipe range 25 ㎝, diameter 2.5 ㎝, lower end absorbent cotton seals, and pipe shaft gauze wraps up 2 layers, and matrix 3 ㎝ is exposed in upper end, is convenient to irrigate.At culture matrix center plantation glutinous rehmannia reed head, from 4, irrigated 500ml soil extraction every 10 days, ensure the normal growth of glutinous rehmannia, now agrafe 7 is closed.According to experimental design, expand early stage, block root at glutinous rehmannia seedling stage, block root elongating stage, block root to expand mid-term, block root and expand later stage and Jiaozhuo, the irrigation soils leaching liquor effluent harvest time south soil water and soil mass ratio by 2:1, shake after 4 hr overnight leave standstill and obtain 1000ml, open agrafe 7, solution is now had to flow out from rubber tube, drain gently with syringe 9, gained solution is root exudates.The timely mistake of the root exudates collected 0.22 μm of miillpore filter, and store under being transferred to 4 DEG C of conditions, adopt HPLC to analyze its coumaric acid, P-hydroxybenzoic acid, vanillic acid, syringic acid and ferulaic acid content.
The foregoing is only preferred embodiment of the present invention, all equalizations done according to the present patent application the scope of the claims change and modify, and all should belong to covering scope of the present invention.
Claims (6)
1. an xerophyte root exudates gathering-device, it is characterized in that, comprise a cultivation bucket, described cultivation bucket bottom is funnel-form, described cultivation bucket bottom surface is communicated with a rubber tube, the described rubber tube other end is connected with syringe, soil vermiculite Mixed culture matrix layer and filtering layer is equipped with from top to bottom successively in described cultivation bucket, the irrigation pipe inserting soil vermiculite Mixed culture matrix layer is laid with in described cultivation bucket, Mixed culture matrix layer upper surface is exposed in described irrigation pipe upper end, described irrigation pipe is pvc pipe, pvc pipe lower end absorbent cotton seals, the one-sided boring of pipe shaft is also enclosed with 2 layers of gauze, matrix 3 ~ 4cm is exposed in irrigation pipe upper end, described filtering layer comprises sandy layer and is positioned at the nonwoven layer below sandy layer.
2. a kind of xerophyte root exudates gathering-device according to claim 1, has liquid stopping clamp in the middle part of described rubber tube.
3. an xerophyte root exudates collection method, utilizes a kind of xerophyte root exudates gathering-device as claimed in claim 1, it is characterized in that, comprise the following steps:
(1) first make root exudates gathering-device, in cultivation bucket, fill filtering layer and soil vermiculite Mixed culture matrix;
(2) irrigation pipe is buried, and planting plants;
(3) in plant incubation, the regular irrigation soils leaching liquor of irrigation pipe is passed through, to guarantee the normal growth of plant;
(4) before collecting root exudates, the leaching liquor slowly pouring into source identical with culture matrix soil from irrigation pipe, to there are flowing out in rubber tube, drains the solution of outflow gently with syringe, extracts solution out and is root exudates;
(5) root exudates step (4) collected is the filtering with microporous membrane of 0.22 μm through diameter, and preserves stand-by under the root exudates after filtration is transferred to 4 DEG C of conditions.
4. a kind of xerophyte root exudates collection method according to claim 3, described leaching liquor presses the water and soil mass ratio of 2:1 by the soil in source identical with culture matrix, shakes 4 hours and obtains after overnight stand.
5. a kind of xerophyte root exudates collection method according to claim 3, described culture matrix is the potpourri of soil and vermiculite, volume ratio is 1:0.5 ~ 1:1.5, filtering layer by clean oven dry nonwoven fabrics and lay and nonwoven fabrics on sand form, sand is 0.5 ~ 3kg, particle diameter 0.5 ~ 1mm, described nonwoven fabrics filling thickness is 3 ~ 8cm, limpid to ensure that root exudates collects liquid.
6. a kind of xerophyte root exudates collection method according to claim 3, described cultivation bucket is organic glass bucket, and organic glass bucket inwall is frosted structure, bottomless, high 20 ~ 50cm, diameter 15 ~ 30cm; Bucket shoulder and bucket stem are upside down funnel shape, bucket shoulder height 3 ~ 8cm, bucket stem 3 ~ 8cm, diameter 2 ~ 4cm, and bucket stem end closes up and is convenient to the installation of rubber tube.
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Cited By (1)
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| CN109765077A (en) * | 2019-02-28 | 2019-05-17 | 中国科学院南京土壤研究所 | A kind of root exudates sampler and sampling system |
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| CN104749000A (en) * | 2015-04-02 | 2015-07-01 | 上海交通大学 | Device and method for extracting helophyte root exudates |
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| CN113514269B (en) * | 2021-06-03 | 2022-03-04 | 海南大学 | Root exudate normal position collection system |
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