CN103238521B - Culture medium and culture method for improving asparagus rooting - Google Patents
Culture medium and culture method for improving asparagus rooting Download PDFInfo
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- CN103238521B CN103238521B CN201310194142.1A CN201310194142A CN103238521B CN 103238521 B CN103238521 B CN 103238521B CN 201310194142 A CN201310194142 A CN 201310194142A CN 103238521 B CN103238521 B CN 103238521B
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Abstract
The invention provides a culture method and a culture medium for improving asparagus rooting. Raw materials of the culture medium comprise raw materials of a basic culture medium, and further comprise calcium pantothenate, biotin, ancymidol, a plant growth regulatory substance and a carbohydrate source, wherein the basic culture medium is selected from a plant tissue culture medium in the prior art; and the plant growth regulatory substance comprises auxin promoting plant tissue cytogenesis or kinetin promoting plant cell division. The method comprises the following steps that: Step 1, asparagus seedlings are shifted into a culture medium I for illumination culture, and root primordium forming is induced; and Step 2, the asparagus seedlings cultured in Step 1 are shifted into a culture medium II for continuous culture until rooting. The culture medium can significantly promote the root primordium forming and the root development. The method adopts a two-step rooting method, and the root development can be promoted significantly.
Description
Technical field
The present invention relates to field of plant tissue culture technique, be specifically related to improve the asparagus substratum of taking root and cultural method.
Background technology
Asparagus (Asparagus officinalis L.) has another name called officinalis, Thallus Gracilariae, is internationally recognized " kings of vegetables ".Conventional cultivation mainly adopts seminal propagation, and the excellent F1 generation seed that production adopts relies on external import always, but expensive.By carrying out tissue culture to asparagus improved seeds, not only can Fast-propagation plant, keep the good character of breeding preferably, and complete male plant can be cultivated, thus increase substantially benefit.But the dioecious biological property of asparagus, determines its sexual hybridization breeding and the breeding breeding cycle is long, efficiency is low, and the rooting of vitro seedling of asparagus is very difficult, significantly limit group training effect.
Technical problem to be solved by this invention is for the deficiencies in the prior art, provides a kind of cost-saving, tissue culture method that effectively can improve asparagus rooting rate.
Summary of the invention
One object of the present invention is to provide a kind of substratum improving asparagus and take root.
Another object of the present invention is to provide a kind of cultural method improving asparagus and take root.
Provided by the invention a kind of for improving the substratum that asparagus takes root, the raw material of this substratum, on the basis of minimum medium raw material, also comprises: calcium pantothenate 0.7 ~ 1.5mg/L, vitamin H 0.01 ~ 0.05mg/L, ancymidol 0.05 ~ 0.15mg/L, plant growth regulating substance 0.05mg/L ~ 0.5mg/L, sugared source 15-35g/L; Described volume refers to the volume of total substratum.Described minimum medium is selected from plant tissue culture media of the prior art, comprises MS substratum, White substratum, B5 medium; Preferred MS substratum.Wherein, in the invention, MS, White, B5 minimum medium is commercially available culture medium dry powder; MS culture medium dry powder consumption is 2 ~ 5g/L, B5 medium dry powder consumption is 1 ~ 5g/L, White culture medium dry powder 15 ~ 30g/L; Preferably, MS culture medium dry powder consumption is 4.74g/L, B5 medium dry powder consumption is 3.21g/L, White culture medium dry powder 21.26g/L." g/L " writing a Chinese character in simplified form for unit " grams per liter " in this invention, is interpreted as: the weight preparing the MS dry powder that one liter of complete substratum contains is 4.74g or B5 medium dry powder weight is 3.21g or White culture medium dry powder 21.26g; " mg/L " writes a Chinese character in simplified form for unit " mg/litre ", is interpreted as: prepare the weight containing materials such as ancymidols in one liter of complete substratum; " liter " take volume as the graduated cylinder of 1 liter is standard capacity.
Described plant growth regulating substance comprises the phytokinin promoting plant tissue cell's generation class growth hormone or promote plant cell division; Described growth hormone comprises: NAA, IAA, IBA, wherein preferred NAA or IBA, and the concentration of NAA is 0.05mg/L to 0.1mg/L, and being preferably 0.1mg/L, IBA concentration is 0.05mg/L to 0.1mg/L, is preferably 0.1mg/L.The phytokinin of described promotion plant cell division is selected from KT, 6-BA.NAA, IAA, 6-BA, KT, IBA mentioned in the present invention are the abbreviation of plant growth regulating substance conventional in plant tissue culture technique, wherein, NAA full name is α-naphthaleneacetic acid, IAA full name is indolylacetic acid, IBA full name is indolebutyric acid, 6-BA full name is 6-benzyl aminoadenine, and KT full name is kinetin.
Described substratum also can add gac or perlite, has the effect of more outstanding hestening rooting rate, when adding gac, needing to add agar and being prepared into solid medium in substratum; When adding perlite, in substratum, be prepared into liquid nutrient medium without the need to adding agar.
In described substratum, sugared source preferably sucrose 25g/L, also can be replaced by other carbohydrates, such as white sugar etc.
Another object of the present invention is to provide a kind of method improving asparagus and take root, and the method comprises the steps:
Step 1: asparagus seedling is proceeded to illumination cultivation in substratum I, induction root restriction is formed;
Step 2: asparagus seedling step 1 cultivated proceeds in substratum II again and continues to cultivate until take root;
The raw material of the substratum I in described step 1 is mainly: minimum medium, calcium pantothenate 0.7 ~ 1.5mg/L, vitamin H 0.01 ~ 0.05mg/L, ancymidol 0.05 ~ 0.15mg/L, growth hormone 0.05mg/L ~ 0.5mg/L, sugared source 15-35g/L; Described growth hormone comprises: NAA, IAA, IBA, wherein preferred NAA, and the concentration of NAA is 0.05mg/L to 0.1mg/L, is preferably 0.1mg/L.
The raw material of the substratum II in described step 2 is mainly: minimum medium, calcium pantothenate 0.7 ~ 1.5mg/L, vitamin H 0.01 ~ 0.05mg/L, ancymidol 0.05 ~ 0.15mg/L, growth hormone or phytokinin 0.05mg/L ~ 0.5mg/L, sugared source 15-35g/L; Wherein, growth hormone is selected from NAA, IAA, IBA, and phytokinin is selected from KT, 6-BA, preferred IBA, and IBA concentration is 0.05mg/L to 0.1mg/L, is preferably 0.1mg/L.
Described minimum medium is selected from plant tissue culture media of the prior art, comprises MS substratum, White substratum, B5 medium; Preferred MS substratum.The formula of substratum is write with reference to " plant tissue culture philosophy and technique " Li Sheng.MS, White, B5 minimum medium is commercially available culture medium dry powder; MS culture medium dry powder consumption is 2 ~ 5g/L, B5 medium dry powder consumption is 1 ~ 5g/L, White culture medium dry powder 15 ~ 30g/L; Involved " g/L " writing a Chinese character in simplified form for unit " grams per liter " in substratum I, II of the present invention, is interpreted as: the weight preparing the MS dry powder that one liter of complete substratum contains is 4.74g or B5 medium dry powder weight is 3.21g or White culture medium dry powder 21.26g; " mg/L " writes a Chinese character in simplified form for unit " mg/litre ", is interpreted as: prepare the weight containing materials such as ancymidols in one liter of complete substratum; " liter " take volume as the graduated cylinder of 1 liter is standard capacity.
Described substratum II also can add gac or perlite, has the effect of more outstanding hestening rooting rate, when adding gac, needing to add agar and being prepared into solid medium in substratum; When adding perlite, be prepared into liquid nutrient medium without the need to adding agar in substratum, described perlite is 600-950g/L, preferred 700-850g/L; Described gac is 0.2-3g/L, preferred 1g/L.
Sugared source preferably sucrose in described substratum I, II, also can be replaced by other carbohydrates, such as white sugar etc.
Further, 20 days are no less than in the time of culture medium culturing in described step 1.
Further, in described step 1, culture condition is: culture temperature is 26 ± 2 DEG C, and relative humidity is 70 ± 5%, intensity of illumination 1500 ~ 2000lx, illumination 9-12 hour/day;
Further, medium pH 4-6 in described step 1, preferably 5.7.
Further, in described step 1, the composition of substratum I is preferred: MS+ calcium pantothenate 1mg/L+ vitamin H 0.01mg/L+ ancymidol 0.1mg/L+NAA0.05 ~ 0.1mg/L+ agar 7g/L+ sucrose 25g/L.
Described, the formula of step 2 substratum II is: MS+ calcium pantothenate 1mg/L+ vitamin H 0.01mg/L+ ancymidol 0.1mg/L+IBA0.05 ~ 0.1mg/L+ gac 1g/L+ agar 7g/L+ sucrose 25g/L or MS+ calcium pantothenate 1mg/L+ vitamin H 0.01mg/L+ ancymidol 0.1mg/L+IBA0.05 ~ 0.1mg/L+ perlite 600-950g/L.
Substratum of the present invention, can promote the formation of root restriction significantly and promote the growth of root.The present invention adopts two step rooting, can promote the growth of root significantly.
Following experimental example is used for further proving effect of the present invention, but can not be interpreted as and understand scope contractility.
MS, White, B5 basic medium dry powder used in experimental example of the present invention, embodiment is purchased from Yu Han bio tech ltd, Shanghai, plant growth regulating substance NAA, IAA, KT, 6-BA, IBA, calcium pantothenate, vitamin H, ancymidol are U.S. Sigma and produce, and gac and perlite are purchased from domestic corporation.In addition, basic medium can select commercially available culture medium dry powder, also can prepare use voluntarily according to following formula voluntarily:
MS culture medium prescription:
White culture medium prescription:
B5 medium is filled a prescription:
The impact of experimental example 1 substratum composition on asparagus seedling rooting rate
1. the kind of substratum and preparation
Root media I and II in this experimental example all with MS substratum for minimum medium, MS substratum uses commercially available culture medium dry powder, and consumption is 4.74g/L.Wherein, adding growth hormone in root media I is NAA0.1mg/L, adds growth hormone IBA0.1mg/L in root media II; And the kind of additional nutrient material calcium pantothenate, vitamin H, ancymidol and concentration as shown in table 1; Finally, the equal additional saccharose 25g/L of all root medias, agar 7g/L, PH are 5.7.Carry out packing after being mixed by the substratum of preparation, splendid attire 40ml substratum in each 120ml triangular flask, seals rear sterilizing pan with sealed membrane and carries out autoclave sterilization, and pressure is 1.1Kg/cm
2, temperature 120 DEG C, sterilizing 15 minutes, naturally cools to the substratum after sterilizing and solidifies rear use, and above-mentioned substratum preparation process and sterilising method are known to the skilled person.
Table 1 adds the root media of different nutriments
2. the root culture of asparagus seedling
Aseptically asparagus seedling individual plant is proceeded to the root culture of carrying out the first stage in first stage root media I; Asparagus individual plant was gone to the root culture of carrying out subordinate phase in root media II after 4 weeks by root culture through the first stage again, then cultivated statistics after 4 weeks and to take root strain number calculate rooting rate; The strain of often kind of culture medium inoculated asparagus seedling 100.
Culture condition is: culture temperature is 26 ± 2 DEG C, and relative humidity is 70 ± 5%, intensity of illumination 1500 ~ 2000lx, illumination 9-12 hour/day.
3. result
Drawn by the table 2 in this experimental example, during interpolation three kinds of nutritive substance vitamin Hs, calcium pantothenate, ancymidol, plant growth regulating substances, the rooting rate of asparagus seedling is the highest at the same time, and reach 64%, and observe in process of the test, root system is more flourishing.And when only adding a kind of nutritive substance in vitamin H, calcium pantothenate, ancymidol, asparagus seedling is not all taken root, when adding any two kinds of nutritive substances, the rooting rate of asparagus seedling is also extremely low, as can be seen here, for must add vitamin H, calcium pantothenate, ancymidol three kinds of nutritive substances and plant growth regulating substance in the substratum of asparagus seedling rooting simultaneously.
Table 2 nutritive substance is on the impact of asparagus seedling rooting
The different minimum medium of experimental example 2 is on the impact of asparagus seedling rooting rate
Two stages that asparagus seedling rooting is cultivated use identical minimum medium, and minimum medium kind is MS substratum, White substratum, B5 medium respectively; First stage root media is: on minimum medium basis, add calcium pantothenate 1mg/L, vitamin H 0.01mg/L, ancymidol 0.1mg/L, NAA0.1mg/L, subordinate phase root media is: on minimum medium basis, add calcium pantothenate 1mg/L, vitamin H 0.01mg/L, ancymidol 0.1mg/L, IBA0.1mg/L.The equal additional saccharose 25g/L of all substratum, agar 7g/L, PH are 5.7.In addition, the preparation of this substratum, sterilizing, culture condition are identical with the method in experimental example 1.
Can be drawn by table 3, when nutritive substance and plant growth regulating substance identical, it is remarkable that the kind of minimum medium affects difference to asparagus seedling rooting, but with MS substratum for preferred culture medium.
Table 3 minimum medium is on the impact of asparagus seedling rooting rate
Experimental example 3
When minimum medium and nutritive substance all identical, in often kind of substratum, add different plant growth regulating substances respectively, to determine the impact of the kind of plant growth regulating substance on asparagus seedling rooting rate.
In this experimental example all root medias all with MS substratum for minimum medium, add calcium pantothenate 1mg/L, vitamin H 0.01mg/L and ancymidol 0.1mg/L tri-kinds of nutritive substances more simultaneously, then often kind of substratum adds different plant growth regulating substances respectively, and concrete kind is see table 4; All substratum dose sucrose 25g/L again, agar 7g/L, adjust pH to be 5.7.In addition, the preparation of this substratum, sterilizing, culture condition are identical with the method in experimental example 1.
Can be drawn by table 4: in substratum 7,8,9, after two stages of root media all use identical growth hormone, the rooting rate of asparagus seedling is lower, and the asparagus seedling rooting rate in all the other substratum is all higher than 41%, as can be seen here, in the process that asparagus seedling rooting is cultivated, be good to add growth hormone in first stage root media, and in subordinate phase root culture, the plant growth regulating substance different from the first stage need be added.
Table 4 plant growth regulating substance is on the impact of asparagus seedling rooting rate
Experimental example 4
By adding the nutritive substance (comprising calcium pantothenate, vitamin H and ancymidol) of different concns to confirm the impact of the concentration of nutritive substance on asparagus seedling rooting rate.
Root media in this experimental example take all MS as minimum medium, and add NAA0.1mg/L in first stage root media, add IBA0.1mg/L in subordinate phase root media, add the nutritive substance of different concns simultaneously, its concentration is in table 5; In addition, the equal additional saccharose 25g/L of all substratum, agar 7g/L, PH are 5.7.In addition, the preparation of this substratum, sterilizing, culture condition are identical with the method in experimental example 1.
From table 5, the preferred concentration range of calcium pantothenate is 0.7 ~ 1.5mg/L, the preferred concentration range of vitamin H is 0.01 ~ 0.05mg/L, the preferred concentration range of ancymidol is 0.05 ~ 0.15mg/L; Further, with when calcium pantothenate concentration 1mg/L, biotin concentration 0.01mg/L, ancymidol concentration 0.1mg/L for the best.
Table 5 nutrient concentrations is on the impact of asparagus seedling rooting
Experimental example 5
By adding NAA and IBA of different concns to confirm the impact of plant-growth regulator on asparagus seedling rooting rate in this experimental example.
Root media in this experimental example is all minimum medium with MS, then adds calcium pantothenate 1mg/L, vitamin H 0.01mg/L, ancymidol 0.1mg/L respectively.Add NAA in first stage root media, in subordinate phase root media, add IBA, but the concentration of NAA and IBA slightly difference in often kind of substratum, refer to table 6.In addition, the equal additional saccharose 25g/L of all substratum, agar 7g/L, PH are 5.7.In addition, the preparation of this substratum, sterilizing, culture condition are identical with the method in experimental example 1.
Can be drawn by table 6: the preferred concentration of NAA is the preferred concentration of 0.05mg/L to 0.1mg/L, IBA is 0.05mg/L to 0.1mg/L, and with NAA0.1mg/L and IBA0.1mg/L for the best.
Table 6NAA and IBA concentration is on the impact of asparagus seedling rooting rate
Experimental example 6
Root media in this experimental example, except interpolation nutritive substance and plant growth regulating substance, separately addition of gac or perlite, to work out the preferred culture medium composition of applicable asparagus seedling rooting.
1. first stage root culture
(1) preparation of first stage root media
Based on MS substratum, add plant-growth regulator NAA0.1mg/L respectively, nutritive substance calcium pantothenate 1mg/L, vitamin H 0.01mg/L, ancymidol 0.1mg/L, then add sucrose 25g/L and agar 7g/L;
Carry out packing after being mixed by the substratum of preparation, the substratum of splendid attire 40ml in each 120ml triangular flask, seals rear sterilizing pan with sealed membrane and carries out autoclave sterilization, and pressure is 1.1Kg/cm
2, temperature 120 DEG C, sterilizing 15 minutes, naturally cools to the substratum after sterilizing and solidifies rear use.
(2) first stage root culture
Aseptically asparagus seedling individual plant is proceeded to the root culture of carrying out the first stage in first stage root media I; Every bottle of 5 strains, totally 20 bottles; Culture condition is: culture temperature is 26 ± 2 DEG C, and relative humidity is 70 ± 5%, intensity of illumination 1500 ~ 2000lx, illumination 9-12 hour/day.
2. the root culture of subordinate phase
(1) preparation of subordinate phase root media
Asparagus individual plant was gone to the root culture of carrying out subordinate phase in root media II after 4 weeks by root culture through the first stage again, wherein, the formula of root media II (1) is: MS+ calcium pantothenate 1mg/L+ vitamin H 0.01mg/L+ ancymidol 0.1mg/L+IBA0.1mg/L+ gac 1g/L+ agar 7g/L+ sucrose 25g/L, and the formula of root media II (2) is: MS+ calcium pantothenate 1mg/L+ vitamin H 0.01mg/L+ ancymidol 0.1mg/L+IBA0.1mg/L+ perlite 750g/L+ sucrose 25g/L; The formula of root media II (3) is: MS+ calcium pantothenate 1mg/L+ vitamin H 0.01mg/L+ ancymidol 0.1mg/L+IBA0.1mg/L+ agar 7g/L+ sucrose 25g/L.
The compound method of root media II (1) is: substratum based on MS, add plant-growth regulator IBA0.05mg/L respectively, nutritive substance calcium pantothenate 1mg/L, vitamin H 0.01mg/L, ancymidol 0.1mg/L, gac 1g/L, then add sucrose 25g/L and agar 7g/L; Carry out packing after being mixed by the substratum of preparation, the substratum of splendid attire 40ml in each 120ml triangular flask, seals rear sterilizing pan with sealed membrane and carries out autoclave sterilization, pressure is 1.1Kg/cm2, temperature 120 DEG C, sterilizing 15 minutes, naturally cools to the substratum after sterilizing and solidifies rear use.
The compound method of root media II (2) is: substratum based on MS, add IBA0.05mg/L respectively, calcium pantothenate 1mg/L, vitamin H 0.01mg/L, ancymidol 0.1mg/L, sucrose 25g/L, stir be mixed with liquid nutrient medium with distilled water constant volume, then in the triangular flask of 120ml, liquid nutrient medium 40ml is loaded respectively, again to each perlite containing adding 30g in the triangular flask of liquid nutrient medium, then seal rear sterilizing pan with sealed membrane and carry out autoclave sterilization, pressure is 1.1Kg/cm2, temperature 120 DEG C, sterilizing 15 minutes, use after the substratum naturally cooling after sterilizing.
Compound method and the root media II (1) of root media II (3) are similar.
(2) root culture of subordinate phase
The root culture of carrying out subordinate phase in root media II is gone to by through the asparagus individual plant of first stage root culture after 4 weeks; Often kind of substratum 20 bottles, every bottle of 5 strains; Culture condition is: culture temperature is 26 ± 2 DEG C, and relative humidity is 70 ± 5%, intensity of illumination 1500 ~ 2000lx, illumination 9-12 hour/day; Add up plant number of taking root after cultivating surrounding, the longest root is long.
3. result
Can be drawn by table 7: in the root media of subordinate phase, add gac or perlite has promoter action to taking root of asparagus seedling, and be good to add perlite, rooting rate reaches 88%.
Table 7 gac or perlite are on the impact of asparagus seedling rooting rate
Embodiment
Before the concrete substratum of preparation, for reach nutritive substance and plant growth regulating substance consumption accurately and easy-to-use, can first it is mixed with certain density mother liquor according to usage quantity and deposit to 4 DEG C of refrigerators for subsequent use, concrete compound method be as follows:
(1) preparing calcium pantothenate concentration is the mother liquor of 1mg/L:
Accurately take calcium pantothenate 100mg with electronic balance, and be dissolved in 50ml distilled water and stir, be then settled to 100ml with the graduated cylinder that volume is 100ml, finally by its splendid attire in the reagent bottle that colourless stopping property is stronger, and it is for subsequent use to leave 4 DEG C of refrigerators in.
(2) preparing biotin concentration is the mother liquor of 0.1mg/L:
Accurately take vitamin H 10mg with electronic balance, and be dissolved in 50ml distilled water and stir, be then settled to 100ml with the graduated cylinder that volume is 100ml, finally by its splendid attire in the reagent bottle that colourless stopping property is stronger, and it is for subsequent use to leave 4 DEG C of refrigerators in.
(3) preparing ancymidol concentration is the mother liquor of 0.1mg/L:
Accurately take ancymidol 10mg with electronic balance, and be dissolved in 50ml distilled water and stir, be then settled to 100ml with the graduated cylinder that volume is 100ml, finally by its splendid attire in the reagent bottle that colourless stopping property is stronger, and it is for subsequent use to leave 4 DEG C of refrigerators in.
(4) preparing NAA concentration is the mother liquor of 0.1mg/L:
Accurately take NAA10mg with electronic balance, and be dissolved in 50ml distilled water and stir, be then settled to 100ml with the graduated cylinder that volume is 100ml, finally by its splendid attire in the reagent bottle that colourless stopping property is stronger, and it is for subsequent use to leave 4 DEG C of refrigerators in.
(5) preparing IBA concentration is the mother liquor of 0.1mg/L:
Accurately take IBA10mg with electronic balance, and be dissolved in 50ml distilled water and stir, be then settled to 100ml with the graduated cylinder that volume is 100ml, finally by its splendid attire in the reagent bottle that colourless stopping property is stronger, and it is for subsequent use to leave 4 DEG C of refrigerators in.
(6) preparing IAA concentration is the mother liquor of 0.1mg/L:
Accurately take IAA10mg with electronic balance, and be dissolved in 50ml distilled water and stir, be then settled to 100ml with the graduated cylinder that volume is 100ml, finally by its splendid attire in the reagent bottle that colourless stopping property is stronger, and it is for subsequent use to leave 4 DEG C of refrigerators in.
(7) preparing 6-BA concentration is the mother liquor of 0.1mg/L:
Accurately take 6-BA10mg with electronic balance, and be dissolved in 50ml distilled water and stir, be then settled to 100ml with the graduated cylinder that volume is 100ml, finally by its splendid attire in the reagent bottle that colourless stopping property is stronger, and it is for subsequent use to leave 4 DEG C of refrigerators in.
In addition, the quantity after the substratum packing of preparing in experimental example of the present invention and embodiment all has surplus, this be for prevent sterilizing or due to other reasons cause substratum lost efficacy for subsequent use to do.
Embodiment 1 promotes the formation substratum of root restriction
Formula: MS culture medium dry powder 4.74g/L+ calcium pantothenate 1mg/L+ vitamin H 0.01mg/L+ ancymidol 0.1mg/L+NAA0.1mg/L+ sucrose 25g/L+ agar 7g/L;
Compound method:
(1) take medium component and mix
First use balance accurate weighing MS culture medium dry powder 4.74g, be placed in 1000ml beaker, and add the stirring of 400ml distilled water; Again with liquid-transfering gun measure the calcium pantothenate of appropriate volume respectively, vitamin H, ancymidol, NAA mother liquor add and above-mentionedly to stir containing in the beaker of MS; Finally, add sucrose 25g, agar 7g mixes.
(2) constant volume
Above-mentioned substratum mother liquor is settled to 1L and stirring and evenly mixing, carries out pH value with NaOH and HCl and be adjusted to 5.7.
(3) packing and sterilizing
Carry out packing after being mixed by the substratum of preparation, the substratum of splendid attire 40ml in each 120ml triangular flask, seals rear sterilizing pan with sealed membrane and carries out autoclave sterilization, pressure is 1.1Kg/cm2, temperature 120 DEG C, sterilizing 15 minutes, naturally cools to the substratum after sterilizing and solidifies rear use.
Embodiment 2 promotes that root restriction forms substratum
Formula: MS culture medium dry powder 4.74g/L+ calcium pantothenate 1mg/L+ vitamin H 0.01mg/L+ ancymidol 0.1mg/L+IAA0.1mg/L+ sucrose 25g/L+ agar 7g/L;
Compound method:
(1) take medium component and mix
First use balance accurate weighing MS culture medium dry powder 4.74g, be placed in 1000ml beaker, and add the stirring of 400ml distilled water; Again with liquid-transfering gun measure the calcium pantothenate of appropriate volume respectively, vitamin H, ancymidol, IAA mother liquor add and above-mentionedly to stir containing in the beaker of MS; Finally, add sucrose 25g, agar 7g mixes.
(2) constant volume
Above-mentioned substratum mother liquor is settled to 1L and stirring and evenly mixing, carries out pH value with NaOH and HCl and be adjusted to 5.7.
(3) packing and sterilizing
Carry out packing after being mixed by the substratum of preparation, the substratum of splendid attire 40ml in each 120ml triangular flask, seals rear sterilizing pan with sealed membrane and carries out autoclave sterilization, pressure is 1.1Kg/cm2, temperature 120 DEG C, sterilizing 15 minutes, naturally cools to the substratum after sterilizing and solidifies rear use.
Embodiment 3 promotes that root forms substratum
Formula: MS culture medium dry powder 4.74g/L+ calcium pantothenate 1mg/L+ vitamin H 0.01mg/L+ ancymidol 0.1mg/L+IBA0.1mg/L+ sucrose 25g/L+ agar 7g/L.
Compound method:
(1) take medium component and mix
First use balance accurate weighing MS culture medium dry powder 4.74g, be placed in 1000ml beaker, and add the stirring of 400ml distilled water; Again with liquid-transfering gun measure the calcium pantothenate of appropriate volume respectively, vitamin H, ancymidol, IBA mother liquor add and above-mentionedly to stir containing in the beaker of MS; Finally, add sucrose 25g, agar 7g mixes.
(2) constant volume
Above-mentioned substratum mother liquor is settled to 1L and stirring and evenly mixing, carries out pH value with NaOH and HCl and be adjusted to 5.7.
(3) packing and sterilizing
Carry out packing after being mixed by the substratum of preparation, the substratum of splendid attire 40ml in each 120ml triangular flask, seals rear sterilizing pan with sealed membrane and carries out autoclave sterilization, pressure is 1.1Kg/cm2, temperature 120 DEG C, sterilizing 15 minutes, naturally cools to the substratum after sterilizing and solidifies rear use.
Embodiment 4 promotes that root forms substratum
Formula:
MS culture medium dry powder 4.74g/L+ calcium pantothenate 1mg/L+ vitamin H 0.01mg/L+ ancymidol 0.1mg/L+IBA0.1mg/L+ sucrose 25g/L+ gac 1g/L+ agar 7g/L;
Compound method is:
(1) take medium component and mix
First use balance accurate weighing MS culture medium dry powder 4.74g, be placed in 1000ml beaker, and add the stirring of 400ml distilled water; Again with liquid-transfering gun measure the calcium pantothenate of appropriate volume respectively, vitamin H, ancymidol, NAA mother liquor add and above-mentionedly to stir containing in the beaker of MS; Finally, add sucrose 25g, agar 7g, gac 1g mix.
(2) constant volume
Above-mentioned substratum mother liquor is settled to 1L and stirring and evenly mixing, carries out pH value with NaOH and HCl and be adjusted to 5.7.
(3) packing and sterilizing
Carry out packing after being mixed by the substratum of preparation, the substratum of splendid attire 40ml in each 120ml triangular flask, seals rear sterilizing pan with sealed membrane and carries out autoclave sterilization, pressure is 1.1Kg/cm2, temperature 120 DEG C, sterilizing 15 minutes, naturally cools to the substratum after sterilizing and solidifies rear use.
Embodiment 5 promotes that root forms substratum
Formula:
MS+ calcium pantothenate 1mg/L+ vitamin H 0.01mg/L+ ancymidol 0.1mg/L+IBA0.1mg/L+ sucrose 25g/L+ perlite 750g/L.
Compound method:
(1) take medium component and mix
First use balance accurate weighing MS culture medium dry powder 4.74g, be placed in 1000ml beaker, and add the stirring of 400ml distilled water; Again with liquid-transfering gun measure the calcium pantothenate of appropriate volume respectively, vitamin H, ancymidol, IBA mother liquor add and above-mentionedly to stir containing in the beaker of MS; Finally, add sucrose 25g to mix.
(2) constant volume
Above-mentioned substratum mother liquor is settled to 1L and stirring and evenly mixing, carries out pH value with NaOH and HCl and be adjusted to 5.7.
(3) packing and sterilizing
Carry out packing after being mixed by the substratum of preparation, the substratum of splendid attire 40ml in each 120ml triangular flask, seals rear sterilizing pan with sealed membrane and carries out autoclave sterilization, and pressure is 1.1Kg/cm
2, temperature 120 DEG C, sterilizing 15 minutes, uses the liquid nutrient medium naturally cooling after sterilizing.
Take separately 750g perlite, carry out autoclave sterilization with after container splendid attire with sterilizing pan, pressure is 1.1Kg/cm
2, temperature 120 DEG C, sterilizing 15 minutes, uses after the perlite naturally cooling of sterilizing.
(4) perlite is added in liquid medium within
On Bechtop, by sterilizing and cooled perlite is distributed in liquid nutrient medium, add perlite 30g in every bottle of substratum, then seal with sealed membrane rear for subsequent use.
Embodiment 6 promotes that root forms substratum
Formula: MS culture medium dry powder 4.74g/L+ calcium pantothenate 1mg/L+ vitamin H 0.01mg/L+ ancymidol 0.1mg/L+6-BA0.1mg/L+ sucrose 25g/L+ agar 7g/L.
Compound method:
(1) take medium component and mix
First use balance accurate weighing MS culture medium dry powder 4.74g, be placed in 1000ml beaker, and add the stirring of 400ml distilled water; Again with liquid-transfering gun measure the calcium pantothenate of appropriate volume respectively, vitamin H, ancymidol, 6-BA mother liquor add and above-mentionedly to stir containing in the beaker of MS; Finally, add sucrose 25g, agar 7g mixes.
(2) constant volume
Above-mentioned substratum mother liquor is settled to 1L and stirring and evenly mixing, carries out pH value with NaOH and HCl and be adjusted to 5.7.
(3) packing and sterilizing
Carry out packing after being mixed by the substratum of preparation, the substratum of splendid attire 40ml in each 120ml triangular flask, seals rear sterilizing pan with sealed membrane and carries out autoclave sterilization, pressure is 1.1Kg/cm2, temperature 120 DEG C, sterilizing 15 minutes, naturally cools to the substratum after sterilizing and solidifies rear use.
Embodiment 7 promotes that root forms substratum
Formula: MS culture medium dry powder 4.74g/L+ calcium pantothenate 1mg/L+ vitamin H 0.01mg/L+ ancymidol 0.1mg/L+IBA0.08mg/L+ sucrose 25g/L+ agar 7g/L.
Compound method:
(1) take medium component and mix
First use balance accurate weighing MS culture medium dry powder 4.74g, be placed in 1000ml beaker, and add the stirring of 400ml distilled water; Again with liquid-transfering gun measure the calcium pantothenate of appropriate volume respectively, vitamin H, ancymidol, IBA mother liquor add and above-mentionedly to stir containing in the beaker of MS; Finally, add sucrose 25g, agar 7g mixes.
(2) constant volume
Above-mentioned substratum mother liquor is settled to 1L and stirring and evenly mixing, carries out pH value with NaOH and HCl and be adjusted to 5.7.
(3) packing and sterilizing
Carry out packing after being mixed by the substratum of preparation, the substratum of splendid attire 40ml in each 120ml triangular flask, seals rear sterilizing pan with sealed membrane and carries out autoclave sterilization, pressure is 1.1Kg/cm2, temperature 120 DEG C, sterilizing 15 minutes, naturally cools to the substratum after sterilizing and solidifies rear use.
The cultural method that embodiment 8 asparagus takes root
Step 1: adventitious bud inducing and multiplication culture:
(1) preparation of adventitious bud induction culture base:
With MS substratum for minimum medium, MS substratum uses commercially available culture medium dry powder 4.74g/L, then adds 6-BA0.8mg/L, NAA0.1mg/L, agar 7g/L, sucrose 25g/L, medium pH 5.7.Carry out packing after being mixed by the substratum of preparation, splendid attire 40ml substratum in each 120ml triangular flask, seals rear sterilizing pan with sealed membrane and carries out autoclave sterilization, pressure is 1.1Kg/cm2, temperature 120 DEG C, sterilizing 15 minutes, naturally cools to the substratum after sterilizing and solidifies rear use.
(2) adventitious bud inducing and multiplication culture
Choose healthy and strong aseptic asparagus seedling, aseptically cut stem apex, be inoculated in asparagus adventitious bud induction culture base and carry out adventitious bud inducing, go to again after 20 days in proliferated culture medium and carry out aseptic seedling and propagating cultivation; Above-mentioned culture condition is: culture temperature is 26 ± 2 DEG C, and relative humidity is 70 ± 5%, intensity of illumination 1500 ~ 2000lx, illumination 9-12 hour/day, medium pH 5.7.
Adventitious bud induction culture based formulas is as follows: MS+6-BA0.8mg/L+NAA0.1mg/L+ agar 7g/L+ sucrose 25g/L; Proliferation culture medium formula is identical with adventitious bud induction culture base.
Step 2: first stage root culture
(1) first stage root media
Root media I is filled a prescription: MS culture medium dry powder 4.74g/L+ calcium pantothenate 1mg/L+ vitamin H 0.01mg/L+ ancymidol 0.1mg/L+NAA0.1mg/L+ agar 7g/L+ sucrose 25g/L.
Carry out packing after being mixed by the substratum of preparation, the substratum of splendid attire 40ml in each 120ml triangular flask, seals rear sterilizing pan with sealed membrane and carries out autoclave sterilization, pressure is 1.1Kg/cm2, temperature 120 DEG C, sterilizing 15 minutes, naturally cools to the substratum after sterilizing and solidifies rear use.
(2) first stage root culture
Aseptically the asparagus individual plant obtained through step 1 is proceeded to the root culture of carrying out the first stage in first stage root media I; Every bottle of 5 strains, totally 20 bottles; Culture condition is: culture temperature is 26 ± 2 DEG C, and relative humidity is 70 ± 5%, intensity of illumination 1500 ~ 2000lx, illumination 9-12 hour/day.
Step 3: the root culture of subordinate phase
(1) subordinate phase root media
Root media II formula is: MS culture medium dry powder 4.74g/L+ calcium pantothenate 1mg/L+ vitamin H 0.01mg/L+ ancymidol 0.1mg/L+IBA0.1mg/L+ gac 1g/L+ agar 7g/L+ sucrose 25g/L.
Carry out packing after being mixed by the substratum of preparation, the substratum of splendid attire 40ml in each 120ml triangular flask, seals rear sterilizing pan with sealed membrane and carries out autoclave sterilization, pressure is 1.1Kg/cm2, temperature 120 DEG C, sterilizing 15 minutes, naturally cools to the substratum after sterilizing and solidifies rear use.
(2) root culture of subordinate phase
The root culture of carrying out subordinate phase in root media II is gone to by through the asparagus individual plant of first stage root culture after 4 weeks; Often kind of substratum 20 bottles, every bottle of 5 strains; Culture condition is: culture temperature is 26 ± 2 DEG C, and relative humidity is 70 ± 5%, intensity of illumination 1500 ~ 2000lx, illumination 9-12 hour/day.
Step 4: rooting rate is added up
Add up rooting rate in the root culture through subordinate phase after 4 weeks, reach 76%, and Plant state is good, suitable booth is transplanted.
The cultural method that embodiment 9 asparagus takes root
Step 1: adventitious bud inducing and multiplication culture:
(1) preparation of adventitious bud induction culture base:
With MS substratum for minimum medium, MS substratum uses commercially available culture medium dry powder 4.74g/L, then adds 6-BA0.8mg/L, NAA0.1mg/L, agar 7g/L, sucrose 25g/L, medium pH 5.7.Carry out packing after being mixed by the substratum of preparation, splendid attire 40ml substratum in each 120ml triangular flask, seals rear sterilizing pan with sealed membrane and carries out autoclave sterilization, pressure is 1.1Kg/cm2, temperature 120 DEG C, sterilizing 15 minutes, naturally cools to the substratum after sterilizing and solidifies rear use.
(2) adventitious bud inducing and multiplication culture
Choose healthy and strong aseptic asparagus seedling, aseptically cut stem apex, be inoculated in asparagus adventitious bud induction culture base and carry out adventitious bud inducing, go to again after 20 days in proliferated culture medium and carry out aseptic seedling and propagating cultivation; Above-mentioned culture condition is: culture temperature is 26 ± 2 DEG C, and relative humidity is 70 ± 5%, intensity of illumination 1500 ~ 2000lx, illumination 9-12 hour/day, medium pH 5.7.
Adventitious bud induction culture based formulas is as follows: MS+6-BA0.8mg/L+NAA0.1mg/L+ agar 7g/L+ sucrose 25g/L; Proliferation culture medium formula is identical with adventitious bud induction culture base.
Step 2: first stage root culture
(1) first stage root media
Root media I is filled a prescription: MS culture medium dry powder 4.74g/L+ calcium pantothenate 1mg/L+ vitamin H 0.01mg/L+ ancymidol 0.1mg/L+NAA0.05mg/L+ agar 7g/L+ sucrose 25g/L.
Carry out packing after being mixed by the substratum of preparation, the substratum of splendid attire 40ml in each 120ml triangular flask, seals rear sterilizing pan with sealed membrane and carries out autoclave sterilization, and pressure is 1.1Kg/cm
2, temperature 120 DEG C, sterilizing 15 minutes, naturally cools to the substratum after sterilizing and solidifies rear use.
(2) first stage root culture
Aseptically the asparagus individual plant obtained through step 1 is proceeded to the root culture of carrying out the first stage in first stage root media I; Every bottle of 5 strains, totally 20 bottles; Culture condition is: culture temperature is 26 ± 2 DEG C, and relative humidity is 70 ± 5%, intensity of illumination 1500 ~ 2000lx, illumination 9-12 hour/day.
Step 3: the root culture of subordinate phase
(1) subordinate phase root media
Root media II formula is: MS culture medium dry powder 4.74g/L+ calcium pantothenate 1mg/L+ vitamin H 0.01mg/L+ ancymidol 0.1mg/L+IBA0.05mg/L+ gac 1g/L+ agar 7g/L+ sucrose 25g/L.
Carry out packing after being mixed by the substratum of preparation, the substratum of splendid attire 40ml in each 120ml triangular flask, seals rear sterilizing pan with sealed membrane and carries out autoclave sterilization, pressure is 1.1Kg/cm2, temperature 120 DEG C, sterilizing 15 minutes, naturally cools to the substratum after sterilizing and solidifies rear use.
(2) root culture of subordinate phase
The root culture of carrying out subordinate phase in root media II is gone to by through the asparagus individual plant of first stage root culture after 4 weeks; Often kind of substratum 20 bottles, every bottle of 5 strains; Culture condition is: culture temperature is 26 ± 2 DEG C, and relative humidity is 70 ± 5%, intensity of illumination 1500 ~ 2000lx, illumination 9-12 hour/day.
Step 4: rooting rate is added up
Add up rooting rate in the root culture through subordinate phase after 4 weeks, reach 70%, and Plant state is good, suitable booth is transplanted.
The cultural method that embodiment 10 asparagus takes root
Step 1: adventitious bud inducing and multiplication culture:
(1) preparation of adventitious bud induction culture base:
With MS substratum for minimum medium, MS substratum uses commercially available culture medium dry powder 4.74g/L, then adds 6-BA0.8mg/L, NAA0.1mg/L, agar 7g/L, sucrose 25g/L, medium pH 5.7.Carry out packing after being mixed by the substratum of preparation, splendid attire 40ml substratum in each 120ml triangular flask, seals rear sterilizing pan with sealed membrane and carries out autoclave sterilization, and pressure is 1.1Kg/cm
2, temperature 120 DEG C, sterilizing 15 minutes, naturally cools to the substratum after sterilizing and solidifies rear use.
(2) adventitious bud inducing and multiplication culture
Choose healthy and strong aseptic asparagus seedling, aseptically cut stem apex, be inoculated in asparagus adventitious bud induction culture base and carry out adventitious bud inducing, go to again after 20 days in proliferated culture medium and carry out aseptic seedling and propagating cultivation; Above-mentioned culture condition is: culture temperature is 26 ± 2 DEG C, and relative humidity is 70 ± 5%, intensity of illumination 1500 ~ 2000lx, illumination 9-12 hour/day, medium pH 5.7.
Adventitious bud induction culture based formulas is as follows: MS+6-BA0.8mg/L+NAA0.1mg/L+ agar 7g/L+ sucrose 25g/L; Proliferation culture medium formula is identical with adventitious bud induction culture base.
Step 2: first stage root culture
(1) first stage root media
Root media I is filled a prescription: MS culture medium dry powder 4.74g/L+ calcium pantothenate 1mg/L+ vitamin H 0.01mg/L+ ancymidol 0.1mg/L+NAA0.05mg/L+ agar 7g/L+ sucrose 25g/L.
Carry out packing after being mixed by the substratum of preparation, the substratum of splendid attire 40ml in each 120ml triangular flask, seals rear sterilizing pan with sealed membrane and carries out autoclave sterilization, pressure is 1.1Kg/cm2, temperature 120 DEG C, sterilizing 15 minutes, naturally cools to the substratum after sterilizing and solidifies rear use.
(2) first stage root culture
Aseptically the asparagus individual plant obtained through step 1 is proceeded to the root culture of carrying out the first stage in first stage root media I; Every bottle of 5 strains, totally 20 bottles; Culture condition is: culture temperature is 26 ± 2 DEG C, and relative humidity is 70 ± 5%, intensity of illumination 1500 ~ 2000lx, illumination 9-12 hour/day.
Step 3: the root culture of subordinate phase
(1) subordinate phase root media
Root media II formula is: MS culture medium dry powder 4.74g/L+ calcium pantothenate 1mg/L+ vitamin H 0.01mg/L+ ancymidol 0.1mg/L+IBA0.05mg/L+ perlite 750g/L+ sucrose 25g/L.
The compound method of root media II as described in Example 5.
(2) root culture of subordinate phase
The root culture of carrying out subordinate phase in root media II is gone to by through the asparagus individual plant of first stage root culture after 4 weeks; Often kind of substratum 20 bottles, every bottle of 5 strains; Culture condition is: culture temperature is 26 ± 2 DEG C, and relative humidity is 70 ± 5%, intensity of illumination 1500 ~ 2000lx, illumination 9-12 hour/day.
Step 4: rooting rate is added up
Add up rooting rate in the root culture through subordinate phase after 4 weeks, reach 84%, and Plant state is good, suitable booth is transplanted.
The cultural method that embodiment 11 asparagus takes root
Step 1: adventitious bud inducing and multiplication culture:
(1) preparation of adventitious bud induction culture base:
With MS substratum for minimum medium, MS substratum uses commercially available culture medium dry powder 4.74g/L, then adds 6-BA0.8mg/L, NAA0.1mg/L, agar 7g/L, sucrose 25g/L, medium pH 5.7.Carry out packing after being mixed by the substratum of preparation, splendid attire 40ml substratum in each 120ml triangular flask, seals rear sterilizing pan with sealed membrane and carries out autoclave sterilization, pressure is 1.1Kg/cm2, temperature 120 DEG C, sterilizing 15 minutes, naturally cools to the substratum after sterilizing and solidifies rear use.
(2) adventitious bud inducing and multiplication culture
Choose healthy and strong aseptic asparagus seedling, aseptically cut stem apex, be inoculated in asparagus adventitious bud induction culture base and carry out adventitious bud inducing, go to again after 20 days in proliferated culture medium and carry out aseptic seedling and propagating cultivation; Above-mentioned culture condition is: culture temperature is 26 ± 2 DEG C, and relative humidity is 70 ± 5%, intensity of illumination 1500 ~ 2000lx, illumination 9-12 hour/day, medium pH 5.7.
Adventitious bud induction culture based formulas is as follows: MS+6-BA0.8mg/L+NAA0.1mg/L+ agar 7g/L+ sucrose 25g/L; Proliferation culture medium formula is identical with adventitious bud induction culture base.
Step 2: first stage root culture
(1) first stage root media
Root media I is filled a prescription: MS culture medium dry powder 4.74g/L+ calcium pantothenate 1mg/L+ vitamin H 0.01mg/L+ ancymidol 0.1mg/L+NAA0.1mg/L+ agar 7g/L+ sucrose 25g/L.
Carry out packing after being mixed by the substratum of preparation, the substratum of splendid attire 40ml in each 120ml triangular flask, seals rear sterilizing pan with sealed membrane and carries out autoclave sterilization, pressure is 1.1Kg/cm2, temperature 120 DEG C, sterilizing 15 minutes, naturally cools to the substratum after sterilizing and solidifies rear use.
(2) first stage root culture
Aseptically the asparagus individual plant obtained through step 1 is proceeded to the root culture of carrying out the first stage in first stage root media I; Every bottle of 5 strains, totally 20 bottles; Culture condition is: culture temperature is 26 ± 2 DEG C, and relative humidity is 70 ± 5%, intensity of illumination 1500 ~ 2000lx, illumination 9-12 hour/day.
Step 3: the root culture of subordinate phase
(1) subordinate phase root media
Root media II formula is: MS culture medium dry powder 4.74g/L+ calcium pantothenate 1mg/L+ vitamin H 0.01mg/L+ ancymidol 0.1mg/L+IBA0.1mg/L+ perlite 750g/L+ sucrose 25g/L.
The compound method of root media II as described in Example 5.
(2) root culture of subordinate phase
The root culture of carrying out subordinate phase in root media II is gone to by through the asparagus individual plant of first stage root culture after 4 weeks; Often kind of substratum 20 bottles, every bottle of 5 strains; Culture condition is: culture temperature is 26 ± 2 DEG C, and relative humidity is 70 ± 5%, intensity of illumination 1500 ~ 2000lx, illumination 9-12 hour/day.
Step 4: rooting rate is added up
Add up rooting rate in the root culture through subordinate phase after 4 weeks, reach 88%, and Plant state is good, suitable booth is transplanted.
Claims (6)
1. one kind for improving the substratum that asparagus takes root, the raw material of this substratum, on the basis of minimum medium raw material, also comprises: calcium pantothenate 0.7 ~ 1.5mg/L, vitamin H 0.01 ~ 0.05mg/L, ancymidol 0.05 ~ 0.15mg/L, plant growth regulating substance 0.05 ~ 0.5mg/L and sugared source 15 ~ 35g/L;
Described plant growth regulating substance comprises the phytokinin promoting plant tissue cell's generation class growth hormone or promote plant cell division;
Described minimum medium is selected from MS substratum, White substratum or B5 medium;
Described growth hormone is selected from NAA, IAA or IBA;
The phytokinin of described promotion plant cell division is selected from KT or 6-BA.
2. substratum as claimed in claim 1, described substratum also adds gac or perlite.
3. substratum as claimed in claim 1, described sugared source is sucrose 25g/L.
4. improve the method that asparagus takes root, the method comprises the steps:
Step 1: asparagus seedling is proceeded to illumination cultivation in substratum I, induction root restriction is formed;
Step 2: asparagus seedling step 1 cultivated proceeds in substratum II again and continues to cultivate until take root;
The raw material of the substratum I in described step 1 is mainly: minimum medium, calcium pantothenate 0.7 ~ 1.5mg/L, vitamin H 0.01 ~ 0.05mg/L, ancymidol 0.05 ~ 0.15mg/L, growth hormone 0.05 ~ 0.1mg/L, sugared source 15-35g/L; Described growth hormone be selected from NAA, IAA, IBA any one;
The raw material of the substratum II in described step 2 is mainly: minimum medium, calcium pantothenate 0.7 ~ 1.5mg/L, vitamin H 0.01 ~ 0.05mg/L, ancymidol 0.05 ~ 0.15mg/L, growth hormone or phytokinin 0.05mg/L ~ 0.1mg/L, sugared source 15-35g/L; Wherein, growth hormone be selected from NAA, IAA, IBA any one, and the growth hormone selected in this step is different from the growth hormone selected in step 1; Phytokinin is selected from KT or 6-BA;
Minimum medium is selected from MS substratum, White substratum or B5 medium; MS culture medium dry powder consumption is 2 ~ 5g/L, B5 medium dry powder consumption is 1 ~ 5g/L, White culture medium dry powder 15 ~ 30g/L.
5. method as claimed in claim 4, described substratum II also adds gac or perlite.
6. method as claimed in claim 4, in described step 1, culture condition is: culture temperature is 26 ± 2 DEG C, and relative humidity is 70 ± 5%, intensity of illumination 1500 ~ 2000lx, illumination 9-12 hour/day.
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| CN116439131A (en) * | 2023-04-03 | 2023-07-18 | 内蒙古库布其沙漠技术研究院 | Tissue culture seedling raising method for sand-fixing plant polygonum sapidum |
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