CN102217498A - Device for preparing solid culture medium supplied by nonuniform nutrients and application method - Google Patents
Device for preparing solid culture medium supplied by nonuniform nutrients and application method Download PDFInfo
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- CN102217498A CN102217498A CN 201110143144 CN201110143144A CN102217498A CN 102217498 A CN102217498 A CN 102217498A CN 201110143144 CN201110143144 CN 201110143144 CN 201110143144 A CN201110143144 A CN 201110143144A CN 102217498 A CN102217498 A CN 102217498A
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Abstract
The invention relates to a device for preparing a solid culture medium supplied by nonuniform nutrients and an application method. The device comprises a culture dish and a division board, wherein the culture dish consists of an upper cover and a bottom cover; a group of clamping grooves are formed in an inner cavity of the bottom cover; the division board is in hermetic insertion with the clamping grooves; and the inner cavity of the bottom cover is partitioned into a plurality of independent spaces by taking the division board as a boundary. The device is easy and convenient to use; ingredients in each partitioned block of the poured culture medium are uniform, and after being solidified, the culture medium can be used without being diffused; nutrients added locally are accurate and high in repeatability; the clamping grooves can prevent culture medium components on the upper part from diffusing and permeating towards a lower layer with moisture at the bottom, and are high in isolating effect; and the device is suitable for supplying a culturing nutrients on specific root section areas of plant root systems selectively and particularly is suitable for performing in-situ nutrient supply or other medicinal treatment on overground parts or a certain specific organ of whole roots of plants, so that the study is closer to the actual condition of plant growth.
Description
Technical field
The invention belongs to vegetable solid culture apparatus technical field, relate to a kind of device and application method that disposes the agar solid culture flat board of non-homogeneous nutrient or material supply.This device is applicable to be needed in Plant Nutrition, plant physiology and the environmental science plant certain organs and root system particular organization section are carried out the research that topical remedy or nutrient are handled in situ.Be specially adapted to inquire into the axial root system plant seedlings to the supply of plant zones of different or lack the response characteristic and the Regulation Mechanism of various nutrients and water-soluble substances.
Background technology
Root system of plant has the nutrient chemotaxis goes after profits and advoids disadvantages to environment with realization, and mostly the distribution in soil of various nutrient or pollutant is uneven.For the nutrient of studying skewness in the soil needs corresponding device thereof to come the simulated soil environment to the influence and the regulation and control of plant growing, can under laboratory condition, realize supplying some material to root system of plant different tissues site specific in situ.In addition, in recent years, some nutrients and other materials enter in the plant corpus by atmospheric sedimentation and then regulating plant growth causes that day by day people pay attention to, and become new research direction.As, plant can absorb the nitrogen of atmospheric sedimentation by blade.These are not quite similar by the process that blade enters its absorption of nutrient, metabolic process and root system of plant absorption metabolism in the plant corpus, and are also different with its inherent regulatory mechanism to the influence that plant growing produces.In order more in depth to study these inherent mechanisms, must develop the nutrient supply of specific laboratory cultures device realization to the plant Different Organs.The model plant arabidopsis is the ideal material of the above mechanism of further investigation, and present stage does not cooperate the arabidopsis seedling to carry out the specific device of non-homogeneous nutrient supply.Common square culture dish is used for above research, need on the monoblock basal medium that configures, cut out the wide ditch of 3-4mm, add high nutrient solution to corresponding agar block surface afterwards, leave standstill to make in 24 hours to add and use (Zhang and Forde, 1998 after solution fully spreads; Zhang
Et al.,1999; Remans
Et al.,2006).This method is complex operation not only, length consuming time, easily microbiological contamination.And the high concentration nutrient that adds is difficult to accurate quantification, is difficult for diffusion evenly, and error is big between repetition.Simultaneously, only isolate different agar blocks, can't stop in the culture block of upper strata water soluble nutrient fully and lose infiltration to underflow with the air ditch.So very need design and develop the culture apparatus that plant Different Organs, different tissues position is carried out the non-homogeneous nutrient supply of original position at arabidopsis.
Summary of the invention
The technical problem that solves: the present invention is directed to above technical problem, design provides a kind of device and methods for using them that is specially adapted to axial root system plant seedlings such as arabidopsis are carried out the local nutrient supply of Different Organs and different tissues position.This device is easy to use, and the layoutprocedure time is short, and each culture block internal component is even, is difficult for diffusion.
Technical scheme: dispose the device of non-homogeneous nutrient supply solid culture medium, comprise culture dish and demarcation strip, described culture dish is made up of loam cake and bottom, the inner chamber of bottom is provided with one group of draw-in groove, demarcation strip and draw-in groove sealing are pegged graft, and are the boundary with the demarcation strip, and the bottom inner chamber is divided into some independently spaces.
Described culture dish is square, and bottom is provided with at least two draw-in grooves that are lower than culture dish bottom height.
Described division board is the glass bar that light was thrown in bottom and both sides, and the thickness of division board equates that with the draw-in groove inner width length and culture dish bottom are isometric, and division board can insert on the culture dish bottom in arbitrary draw-in groove, works to isolate medium.
Dispose the application method of the device of non-homogeneous nutrient supply solid culture medium, step is: according to the kind quantity n of required medium, choosing n-1 piece division board inserts in the draw-in groove, each component medium is mixed in the culture dish bottom space of directly pouring corresponding site into, after treating the complete cooled and solidified of medium, extract demarcation strip and get final product.Demarcation strip can reuse.
Beneficial effect: apparatus of the present invention are easy to use quick; Each piecemeal internal component of the medium of pouring out is even, and culture medium solidifying can be used, and need not diffusion; Local amount of nutrients of adding is accurate, easily repeats; Draw-in groove can stop in the bottom top nutrient media components with moisture to lower floor's scattering and permeating, isolation effect is good; This device is not only applicable to optionally nutrient supply cultivation is carried out in the special root segment of root system of plant zone, more be applicable to plant shoot or the complete a certain certain organs of root are carried out the nutrient supply or the other drug processing of original position, make research more press close to the actual conditions of plant growing.
Description of drawings
The present invention is further described below in conjunction with drawings and Examples.
Fig. 1: the assembling sectional view of this device, 1 culture dish loam cake, 2 culture dish bottoms among the figure, 3 draw-in grooves, 4 demarcation strips.
Fig. 2: the vertical view of this device bottom, 2 culture dish bottoms among the figure, 3 draw-in grooves.
Fig. 3: with contrive equipment configuration overground part nutrient supply medium (A), underground part nutrient supply medium (B), maturation zone nutrient supply medium (C) and lateral root root division medium (D);
Figure: 4: each culture block internal component of non-homogeneous nutrient supply medium of apparatus of the present invention configuration is even.Wherein, the medium that configures with this device of A; The non-homogeneous nutrient supply medium of B previous methods preparation; The non-homogeneous medium of C previous methods preparation leaves standstill diffusion 2 d; The enlarged image of white arrow indication non-uniform areas among the D C figure.
Fig. 5: the diffusion permeability that apparatus of the present invention dispose local nutrient supply culture plate detects.Wherein, A, B are for before beginning vertically to put into hot-house culture; C, D vertically cultivate 1 d for putting into the greenhouse; E, F are for vertically putting into hot-house culture 5 d; A, C, E are the non-homogeneous nutrient medium of new device configuration; B, D, F are respectively the enlarged images in red arrow indication zone among A, C, the E figure.
Fig. 6: use this device research arabidopsis seedling supplies ammonium for ammonium and root to overground part response.Wherein, A is that overground part is for the influence of ammonium to the arabidopsis seedling; B is that underground part is for the influence of ammonium to the arabidopsis seedling.
Fig. 7: use this device research arabidopsis seedling to the root tip of a root and ripe response for ammonium.
Figure: 8: use this device research arabidopsis seedling to of the response of lateral root root division for ammonium.Wherein, the one-sided lateral root of A is for the influence of ammonium to arabidopsis lateral root length; The one-sided lateral root of B is for the influence of ammonium to arabidopsis secondary lateral root length.
Embodiment
Embodiment 1:
Apparatus of the present invention as shown in Figure 1, during use according to the kind quantity n of required medium, choose n-1 piece division board and insert in the draw-in groove, each component medium is mixed in the culture dish bottom space of directly pouring corresponding site into, treat to extract demarcation strip after the medium cooled and solidified of various piece.
As shown in Figure 3, choose one of them draw-in groove insertion demarcation strip culture dish is divided into two parts up and down, pour high ammonium medium on top into, the bottom is poured normal medium into can be made into overground part for the ammonium medium.Otherwise, can be made into the medium that is used for full root confession ammonium and tip of a root confession ammonium.Get two demarcation strips and insert in two draw-in grooves of about 2 cm, culture dish is divided into upper, middle and lower three parts, pour high ammonium medium at the middle part into, pour normal medium into for two ones up and down, can be made into the root maturation zone for the ammonium medium.Get that two demarcation strips are anti-to insert that distance two draw-in grooves far away are interior not to be divided into left, center, right three parts with culture dish, about two ones pour high ammonium medium into, normal medium is poured at the middle part into can make lateral root root division culture plate (seeing example 2,3 for details).
Embodiment 2:
Local nutrient supply medium with configuration overground part supply high concentration ammonium nutrient is an example, and the interpolation thymol blue makes it present blueness to be different from the culture block of normal components in containing the high nutrient medium of high concentration ammonium ion.As can be seen from Figure 4, very even with its high nutrient culture block internal component of medium of this contrive equipment configuration, and do not need to leave standstill diffusion, and can use after the condensation, save the plenty of time.Simultaneously, because the component of each culture block of medium is just to add and mixing by liquid fraction before a kind of rhyme scheme in Chinese operas serving as the prelude to a complete score for voices, therefore, its high nutrient or content of material add accurately, favorable reproducibility.
The culture plate simulating plant condition of culture that configures is vertically put into hot-house culture, as can be seen from Figure 5, culture plate with this device configuration, owing to have lower draw-in groove to separate medium in the bottom between its inner each culture block, the culture block component on top can not get the culture block scattering and permeating to the bottom with moisture etc.Even place 20 d, still do not find the visible diffusion of bore hole.Solved the scattering and permeating problem that exists when studying local nutrient supply well, kept the stable of each culture block component, thereby the experimental result that obtains is objective reality more.
Embodiment 3:
Most nutrient ions and other materials both can absorb to enter in the plant corpus and also can enter in the plant corpus by the overground part blade by root, particularly ammonium nitrogen.The external source ammonium that the plant Different Organs absorbs arrives the different parts in the plant corpus, participate in different metabolic processes, whether they are the same to the influence of growth and development of plants so, is plant corpus the external source ammonium that absorbs by identical molecule physiological pathway response Different Organs? in order to solve the method that similar query need absorb the external source nutrient in the contact of laboratory simulation plant Different Organs.Can realize this simulation easily with apparatus of the present invention.With the ammonium is the example explanation.1) overground part promptly arabidopsis seedling overground part is comprised leaf and stem for ammonium, and root does not touch high ammonium for ammonium.2) root be meant to supply ammonium to arabidopsis shoot root portion is whole, and overground part does not contact high ammonium for ammonium.Prepare corresponding medium according to example 1 is described.Selecting on normal medium the seedling of good growth 5 d left and right sides growing way unanimities transplants on culture plate as requested and gets final product.After having moved seedling, cover the culture dish lid, with sealing after film seals, vertically place growth room's growth 5 d after, sample analysis.Growth room's condition of culture is, the 16:8 h photoperiod, temperature is 23 ± 1 ℃, and fluorescent lamp intensity is about 100 umol m
-2s
-1
The arabidopsis seedling is well-grown on the medium of preparing out with new method, and reappearance is good between individuality.Show that this culture apparatus itself does not have influence to growth of seedling, can be used for scientific research.From this routine result, overground part has very big-difference for ammonium to the influence of arabidopsis seedling for ammonium and root.Though overground part has all suppressed the growth (Fig. 6) of root system significantly for ammonium for ammonium and root, but overground part mainly is the length (Fig. 6 A) that reduces the quantity of lateral root and then reduce lateral root, and root does not suppress the quantity of lateral root for ammonium but suppress the elongation (Fig. 6 B) of lateral root.Overground part supplies ammonium will be weaker than underground part for ammonium (Fig. 6 A﹠amp to the inhibition strength of main root length; B).The Different Effects that this overground part and root produce root system of plant for ammonium can't be found and be studied with overall broth incubation in the past.Two kinds of different ammonium mode differences on phenotype that supply are hinting regulatory pathway and molecular mechanism that they are inner different probably.We use new equipment and have proposed overground part nutrient material first to the plant study on regulation thinking of root system development particularly.The invention of new equipment provides possibility for similar exploratory development.
Embodiment 4:
Root system of plant is divided into a plurality of sections of organizing according to its function difference, and different sections all may have different to the response sensitivity of environment with feature.Therefore, be necessary the different sections of root are carried out specificity nutrient or other drug processing.Can realize quickly and easily the arabidopsis seedling tip of a root or ripe two specific regions are carried out local nutrient or other material supplies processing with apparatus of the present invention.1) tip of a root promptly has only the high ammonium of tip of a root front end 2 those sections of mm zone contact for ammonium, and other sections of overground part and root are affixed on normal media surface; 2) maturation zone is meant that for ammonium the abapical root segment that digs up the roots all is affixed on high ammonium media surface, and the tip of a root and overground part are affixed on the normal medium on bottom and top respectively.Prepare required culture plate according to example 1, transplanting seedlings as requested gets final product.
Hot-house culture can be found after a couple of days, the tip of a root significantly suppresses root growth (Fig. 7) for ammonium, growth does not suppress to main root but the maturation zone is for ammonium, by being implemented the root regional area, the arabidopsis seedling can find for ammonium, high ammonium to the inhibition of root growth be a kind of must be by the local effect of tip of a root mediation, the tip of a root is that root system is experienced the main site that high ammonium suppresses.
Embodiment 5:
Lateral root is not only the important part of root system of plant, and,, lateral root unlike main root, determines, thereby to have stronger environment plasticity jointly because growing fully by the postembryonal development decision by proembryo and postembryonal development stage.The research lateral root is very necessary to the response characteristic of uneven distribution nutrient.Apparatus of the present invention can realize that the different lateral roots to same young plant carry out the supply experiment of both sides Different Nutrients concentration, and promptly the lateral root root division is for ammonium.Be meant that first, second suitable lateral root of length is carried out the high ammonium of a side to be handled.By the example 1 described lateral root root division culture plate that configures, to on normal medium, the main root section of arabidopsis seedling below the visible lateral root of second of growth 6 d all deduct earlier, move on the fresh normal medium and to recover choosing two suitable seedling of lateral root length that stay after growth 3 d and move on the lateral root root division culture plate, make two lateral roots place the air ditch to separate on the medium of the left and right sides respectively, overground part does not contact with high ammonium medium.5 d grow in vertical placement greenhouse.
Analysis result can find that the lateral root length that contacts high ammonium still is subjected to obvious inhibition, and compared with the control, its relative length has descended 62.0%.And not contacting the heteropleural part of high ammonium, its lateral root length and contrast be difference (Fig. 8) not.This result also proved once more the high ammonium of root to the inhibition of lateral root length need lateral root particularly the lateral root tip of a root directly contact with high ammonium.Directly the high nutrient of the high ammonium medium of contact is cultivated regional secondary lateral root length and is subjected to extremely significantly suppressing, and the normal nutrient that does not contact high ammonium medium is cultivated regional secondary lateral root length and significantly increased, and facilitation is obvious.By example as can be seen, this new equipment can be realized the nutrient ion to medium different blocks supply variable concentrations and kind easily, the very suitable influence that plant lateral roots is formed and grows in order to research specific region supply nutrient ion or other materials.
More than implement example proof apparatus of the present invention and can configure the non-homogeneous nutrient supply culture plate of plant quickly, each culture block inside of the culture plate that configures is uniform and stable, not only be fit to certain organs is carried out each species specificity nutrient and other material supplies processing of original position, also be fit to research root a certain particular organization section or regional various nutrient or material supply the regulation mechanism that influences plant growing.
Claims (4)
1. dispose the device of non-homogeneous nutrient supply solid culture medium, it is characterized in that comprising culture dish and demarcation strip (4), described culture dish is made up of loam cake (1) and bottom (2), the inner chamber of bottom (2) is provided with one group of draw-in groove (3), demarcation strip and draw-in groove sealing are pegged graft, with the demarcation strip is the boundary, the bottom inner chamber can be divided into some independently spaces.
2. the device of the non-homogeneous nutrient supply solid culture of configuration according to claim 1 plate is characterized in that described culture dish is square, and bottom is provided with at least two draw-in grooves that are lower than culture dish bottom height.
3. the device of the non-homogeneous nutrient supply solid culture medium of configuration according to claim 1, it is characterized in that described demarcation strip (4) is the glass bar that light was thrown in bottom and both sides, and the thickness of demarcation strip equates with the draw-in groove inner width, length and culture dish bottom are isometric, demarcation strip can insert on the culture dish bottom in arbitrary draw-in groove, works to isolate medium.
4. dispose the application method of the device of non-homogeneous nutrient supply solid culture medium, it is characterized in that step is: according to the kind quantity n of required medium, choosing n-1 piece division board inserts in the draw-in groove, each component medium is mixed in the culture dish bottom space of directly pouring corresponding site into, extract demarcation strip after solidifying and get final product.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104792750A (en) * | 2015-03-27 | 2015-07-22 | 南方医科大学 | Construction method of cell polarity model |
| CN106212252A (en) * | 2016-09-13 | 2016-12-14 | 谢海燕 | A kind of vertical culture vessel |
| CN110367077A (en) * | 2019-07-08 | 2019-10-25 | 佛山科学技术学院 | A kind of preparation method of multiphase solid culture medium |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| JPS63137671A (en) * | 1986-11-28 | 1988-06-09 | Erumetsukusu:Kk | Petri dish for holding medium |
| CN2322972Y (en) * | 1997-12-04 | 1999-06-09 | 中国科学院新疆化学研究所 | Common culture utensil for plurality of cultures |
| CN201634677U (en) * | 2010-02-27 | 2010-11-17 | 张微 | Novel bacterial culture dish |
| CN201695031U (en) * | 2010-05-17 | 2011-01-05 | 复旦大学 | Partitioned culture vessel with glass bottom |
-
2011
- 2011-05-30 CN CN 201110143144 patent/CN102217498A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS63137671A (en) * | 1986-11-28 | 1988-06-09 | Erumetsukusu:Kk | Petri dish for holding medium |
| CN2322972Y (en) * | 1997-12-04 | 1999-06-09 | 中国科学院新疆化学研究所 | Common culture utensil for plurality of cultures |
| CN201634677U (en) * | 2010-02-27 | 2010-11-17 | 张微 | Novel bacterial culture dish |
| CN201695031U (en) * | 2010-05-17 | 2011-01-05 | 复旦大学 | Partitioned culture vessel with glass bottom |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104792750A (en) * | 2015-03-27 | 2015-07-22 | 南方医科大学 | Construction method of cell polarity model |
| CN104792750B (en) * | 2015-03-27 | 2017-11-28 | 南方医科大学 | The construction method of cell polarity model |
| CN106212252A (en) * | 2016-09-13 | 2016-12-14 | 谢海燕 | A kind of vertical culture vessel |
| CN110367077A (en) * | 2019-07-08 | 2019-10-25 | 佛山科学技术学院 | A kind of preparation method of multiphase solid culture medium |
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Application publication date: 20111019 |