CN103477985B - A kind of regeneration culture medium and cultural method improving echinacea purpurea explant regeneration indefinite bud - Google Patents
A kind of regeneration culture medium and cultural method improving echinacea purpurea explant regeneration indefinite bud Download PDFInfo
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Abstract
The invention belongs to plant biotechnology field.The present invention specifically discloses a kind of regeneration culture medium and the cultural method that improve echinacea purpurea explant regeneration indefinite bud.Described regeneration culture medium is to the difference of the susceptibility of diethylamino ethanol caproate (DA-6) according to the echinacea purpurea explant of Different Ploidy and different sources, in MS formula except adding 15 ~ 60g/L sucrose, 3 ~ 9g/L agar, 0.1 ~ 1.5mg/L BA and 0.01 ~ 0.2mg/L NAA, also add the DA-6 of respective concentration especially.The effect that described regeneration culture medium impels BA evoking adventive bud to regenerate by interpolation DA-6 been significantly enhanced.The present invention effectively can overcome different genotype restricted problem in echinacea purpurea regeneration cultivation, significantly improves adventitious shoot regeneration efficiency, promotes carrying out of echinacea purpurea biotechnology breeding related scientific research work.
Description
Technical field
The present invention relates to plant biotechnology field, particularly, relate to a kind of regeneration culture medium and the cultural method that improve echinacea purpurea explant regeneration indefinite bud.
Background technology
Echinacea purpurea (
echinacea purpureal.) be composite family per nnial herb, have good immunoregulation effect, its extract is a kind of immunopotentiating agent and the immunomodulator that are subject to most attention in recent years in the world.Due to the efficacy effect that it is definite, echinacea purpurea has become a kind of world-renowned medicinal plant.People, in order to obtain the echinacea purpurea of multiple kind, have attempted being applied thereon by multiple animal nutrition, as transgenosis, anther culture, chromosome doubling.Reaching of these objects, all has to pass through a common step, i.e. Induced cultures, produces indefinite bud.Although existing a lot of research launches for echinacea purpurea adventitious shoot regeneration in prior art, its regeneration efficiency is unsatisfactory.
In prior art, when echinacea purpurea explant induction adventitious shoot regeneration, usual employing is containing 15 ~ 60 g/L sucrose, 0.1 ~ 1.5 mg/L BA and 0 ~ 0.15 mg/L NAA, with the MS substratum that 3 ~ 9 g/L are agar solidified, under the culture condition of this regeneration culture medium, the adventitious shoot regeneration efficiency variance of different genotype echinacea purpurea explant is very large, except partial genotype can obtain more suitable regeneration efficiency, the regeneration efficiency of all the other lots of genes types is very low, the quality of sprout is also poor, seriously constrain echinacea purpurea transgenosis, anther culture, carrying out of the Plant Biotechnology researchs such as chromosome doubling, limit the carrying out of echinacea purpurea biotechnology breeding work.
Diethylamino ethanol caproate (DA-6) is a plant growth regulators, is generally used for field cultivation, is rarely used in tissue culture.But also nobody attempts using it for echinacea purpurea and organizing cultivation aspect.
Summary of the invention
When the present invention is in order to overcome echinacea purpurea explant regeneration indefinite bud in prior art, the defect of low, the regeneration bud weight difference of regeneration rate, provides a kind of regeneration culture medium improving echinacea purpurea explant regeneration indefinite bud.
Another object of the present invention is to provide a kind of method improving echinacea purpurea explant regeneration indefinite bud.
The present invention is achieved above-mentioned purpose by the following technical programs:
A kind of regeneration culture medium improving echinacea purpurea explant regeneration indefinite bud, it is characterized in that, regeneration culture medium needed for echinacea purpurea explant except adding the NAA of 15 ~ 60 g/L sucrose, 3 ~ 9 g/L agar, 0.1 ~ 1.5 mg/L BA and 0.01 ~ 0.15 mg/L, also with the addition of diethylamino ethanol caproate (DA-6) especially in MS substratum.
Described echinacea purpurea explant is divided into diploid, triploid, tetraploid or hexaploid echinacea purpurea blade, petiole or root explant according to the difference in ploidy and source.
The amount of the DA-6 added in diploid or the regeneration culture medium needed for triploid echinacea purpurea leaf explant is 0.01 ~ 0.32 mg/L; The amount of the DA-6 added in tetraploid or the regeneration culture medium needed for hexaploid echinacea purpurea leaf explant is 0.01 ~ 0.16 mg/L;
The amount of the DA-6 added in diploid or the regeneration culture medium needed for triploid echinacea purpurea petiole explant is 0.01 ~ 0.16 mg/L; The amount of the DA-6 added in the regeneration culture medium needed for tetraploid echinacea purpurea petiole explant is 0.01 mg/L; The amount of the DA-6 added in the regeneration culture medium needed for hexaploid echinacea purpurea petiole explant is 0.08 ~ 0.16 mg/L;
The amount of the DA-6 added in diploid or the regeneration culture medium needed for triploid echinacea purpurea root explant is 0.01 mg/L; The amount of the DA-6 added in the regeneration culture medium needed for tetraploid echinacea purpurea root explant is 0.01 ~ 0.16 mg/L.
Preferably, the amount of the DA-6 added in described diploid or the regeneration culture medium needed for triploid echinacea purpurea leaf explant is 0.08 ~ 0.16 mg/L; More preferably, the amount of the DA-6 added in described diploid or the regeneration culture medium needed for triploid echinacea purpurea leaf explant is 0.16 mg/L.The amount of the DA-6 added in described tetraploid or the regeneration culture medium needed for hexaploid echinacea purpurea leaf explant is 0.08 ~ 0.16 mg/L, more preferably, the amount of the DA-6 added in tetraploid or the regeneration culture medium needed for hexaploid echinacea purpurea leaf explant is 0.08 mg/L.
Preferably, the amount of the DA-6 added in described diploid or the regeneration culture medium needed for triploid echinacea purpurea petiole explant is 0.01 ~ 0.08 mg/L; More preferably, the amount of the DA-6 added in the regeneration culture medium needed for described diploid echinacea purpurea petiole explant is 0.08 mg/L; The amount of the DA-6 added in the regeneration culture medium needed for described triploid echinacea purpurea petiole explant is 0.01 mg/L.Preferably, the amount of the DA-6 added in the regeneration culture medium needed for described hexaploid echinacea purpurea petiole explant is 0.16 mg/L.
Preferably, the amount of the DA-6 added in the regeneration culture medium needed for described tetraploid echinacea purpurea root explant is 0.01 ~ 0.08mg/L; More preferably, the amount of the DA-6 added in the regeneration culture medium needed for described tetraploid echinacea purpurea root explant is 0.01 mg/L.
A kind of cultural method improving echinacea purpurea explant regeneration indefinite bud, the needs of the component of the regeneration culture medium needed for the echinacea purpurea explant in Different Ploidy described above and source, dissimilar echinacea purpurea explant is seeded in regeneration culture medium suitable separately, it is 25 ± 4 DEG C in culture temperature, intensity of illumination is 1000 ~ 2500 lx, light application time is 10 ~ 16 h/d, condition under cultivate 18 ~ 60 d.The mode of described inoculation is traverse mode, even if the horizontal surface of the square section of petiole explant and substratum is perpendicular.
Diethylamino ethanol caproate (DA-6) is a plant growth regulators, is generally used for field cultivation, is rarely used in tissue culture.But also nobody attempts using it for echinacea purpurea and organizing cultivation aspect.When contriver carries out micropropagation to some Special germplasm material (echinacea purpurea of various Different Ploidy), find that culture is very responsive to adding of DA-6, and at certain concentrations, the regeneration efficiency of indefinite bud significantly improves.So the present invention mainly have studied DA-6 affect related experiment to echinacea purpurea adventitious shoot regeneration.
The present invention has investigated and has added diethylamino ethanol caproate (DA-6) in the medium to the impact of echinacea purpurea explant adventitious shoot regeneration.Present invention uses the explant in 3 kinds of sources: blade, petiole and root explant.Wherein needed for leaf explant, best DA-6 concentration is the highest.The concentration that root explant needs is minimum.The explant of four kinds of Different Ploidy is tested, and the explant of Hyperploidy is more responsive to DA-6: the DA-6 concentration that the explant of Hypoploidy needs is higher.To the DA-6 concentration that the explant of Hypoploidy is applicable to, Hyperploidy explant may be suppressed to produce indefinite bud.Result display of the present invention, appropriately use DA-6, the regeneration of the indefinite bud of echinacea purpurea different explants can significantly promote.
Beneficial effect of the present invention: the invention solves usual echinacea purpurea adventitious shoot regeneration cultivate in the not high and imperfect outstanding problem of regeneration bud quality of regeneration efficiency, for echinacea purpurea biotechnology research particularly for can not produce seed but have that higher plantation is worth three, four, the seedling clone of hexaploid has important can being direct appliedly worth.
figure of description
Fig. 1. different DA-6 concentration is on the impact of diploid explant regeneration indefinite bud; 1 ~ 5 is the diploid leaf explant of DA-6 process with 0,0.01,0.08,0.16,0.32 mg/L respectively; 6 ~ 10 is the diploid petiole explant of DA-6 process with 0,0.01,0.08,0.16,0.32 mg/L respectively; 11 ~ 15 is the diploid root explant of DA-6 process with 0,0.01,0.08,0.16,0.32 mg/L respectively.
Fig. 2. different DA-6 concentration is on the impact of triploid explant regeneration indefinite bud; 1 ~ 5 is the triploid leaf explant of DA-6 process with 0,0.01,0.08,0.16,0.32 mg/L respectively; 6 ~ 10 is the triploid petiole explant of DA-6 process with 0,0.01,0.08,0.16,0.32 mg/L respectively; 11 ~ 15 is the triploid root explant of DA-6 process with 0,0.01,0.08,0.16,0.32 mg/L respectively.
Fig. 3. different DA-6 concentration is on the impact of tetraploid explant regeneration indefinite bud; 1 ~ 5 is the tetraploid leaf explant of DA-6 process with 0,0.01,0.08,0.16,0.32 mg/L respectively; 6 ~ 10 is the tetraploid petiole explant of DA-6 process with 0,0.01,0.08,0.16,0.32 mg/L respectively; 11 ~ 15 is the tetraploid root explant of DA-6 process with 0,0.01,0.08,0.16,0.32 mg/L respectively.
Fig. 4. different DA-6 concentration is on the impact of hexaploid explant regeneration indefinite bud; 1 ~ 5 is the hexaploid leaf explant of DA-6 process with 0,0.01,0.08,0.16,0.32 mg/L respectively; 6 ~ 10 is the hexaploid petiole explant of DA-6 process with 0,0.01,0.08,0.16,0.32 mg/L respectively; 11 ~ 15 is the hexaploid root explant of DA-6 process with 0,0.01,0.08,0.16,0.32 mg/L respectively.
Embodiment
Further describe the present invention below in conjunction with the drawings and specific embodiments, help is understood the present invention by embodiment better, but the present invention is not limited only to following embodiment.Unless stated otherwise, the reagent adopted in embodiment and method are reagent and the method for this area routine use.
Vegetable material: diploid of the present invention, tetraploid echinacea purpurea are that contriver passes through to study acquisition in the early time; The preparation method of diploid, tetraploid echinacea purpurea can with reference to Nilanthi D, Chen X L, Zhao F C, et al. Induction of tetraploids from petiole explants through colchicine treatments in Echinacea purpurea L [J]. BioMed Research International, 2009,2009..Triploid echinacea purpurea is obtained Diploid and Tetraploid hybridization.Hexaploid the process of triploid explant colchicine is doubled rear evoking adventive bud to obtain (process of triploid explant colchicine doubled rear evoking adventive bud obtain the method for hexaploid echinacea purpurea can with reference to this area routine techniques).The ploidy of these plant is identified by root tip chromosomes counting technology all.The aseptic echinacea purpurea seedling of all above materials is all containing in 30 g/L sucrose, 0.01 mg/L NAA, the MS substratum agar solidified with 3 ~ 9 g/L, it is 25 ± 4 DEG C in culture temperature, intensity of illumination is 1000 ~ 2500 lx, and light application time is cultivate under the condition of 10 ~ 16 h/d.
Embodiment 1:
S1. explant prepare: with scissors from group training complete echinacea purpurea seedling clip blade, petiole and Gen Ge several.The blade of Different Ploidy, petiole and root train complete aseptic seedling from the group of Different Ploidy.After tap water institute picking leaves sheet, petiole and root 30 min, Bechtop is placed in all blades, petiole and root the glass beaker that a capacity is 1000 mL, 75% ethanolic soln is added to blade, petiole and root that submergence is all in this beaker, after 1 min, outwell ethanolic soln, retain blade, petiole and root; In this beaker, add 2% chlorine bleach liquor again to blade, petiole and root that submergence is all, after 20 min, outwell chlorine bleach liquor, retain blade, petiole and root; In this beaker, add sterile distilled water again to blade, petiole and root that submergence is all, rinsing blade, petiole and root 2 after min, outwells distilled water, retains blade, petiole and root.After rinsed with sterile water blade, petiole and root 5 times, with sterilized tweezers, all blades, petiole and root are placed on aseptic thieving paper, after blade, petiole and root surface drying, with sterilized scalpel, blade, petiole and root are cut, cut by following specification: blade area: 0.6 cm
2, petiole, root length: 0.8 cm, obtain blade, petiole and root explant.
S2. diplontic echinacea purpurea blade, petiole and root explant are inoculated into respectively containing different concns DA-6(0,0.01,0.08,0.16,0.32 mg/L) regeneration culture medium on (regeneration culture medium is in MS substratum, add 30 g/L sucrose, 4.5 g/L agar, 0.3 mg/L BA and 0.01 mg/L NAA).The situation of test adventitious bud inducing regeneration.
Above-mentioned containing different concns DA-6(0,0.01,0.08,0.16,0.32 mg/L) regeneration culture medium substratum collocation method be specially: first in MS substratum, add 30 g/L sucrose, 4.5 g/L agar, 0.3 mg/L BA and 0.01 mg/L NAA, after above composition is configured, autoclaving, when substratum room temperature is cooled to non-scald on hand, in aseptic operating platform, add the DA-6 of each concentration configured with sterilized water, make the final concentration of DA-6 in regeneration culture medium for (0,0.01,0.08,0.16 or 0.32 mg/L).
The culture condition of adventitious bud inducing regeneration is be seeded in by dissimilar echinacea purpurea explant in regeneration culture medium suitable separately, it is 25 ± 4 DEG C in culture temperature, intensity of illumination is 1000 ~ 2500 lx, and light application time is cultivate 35 d under the condition of 10 ~ 16 h/d.
From table 1 and Fig. 1, the DA-6 being applicable to concentration significantly can promote the adventitious bud induction frequency of whole 3 kinds of explants.The DA-6 concentration (0.16 mg/L) that blade obtains best inductivity is higher than petiole and root.And when concentration is up to 0.32 mg/L, the efficiency that leaf explant induction produces indefinite bud is not suppressed yet.Under this concentration, (0.32 mg/L) has created the suppression of 22% to petiole explant, and root explant does not produce indefinite bud substantially.
Table 1 different concns DA-6 is on the impact of diploid explant regeneration indefinite bud
Note: the numerical value of each row different letter representation below significant difference (P<0.05) in Duncan test in table.
Embodiment 2
S1. the preparation of explant: with embodiment 1.
S2 triploid echinacea purpurea blade, petiole and root explant are inoculated into respectively containing different concns DA-6(0,0.01,0.08,0.16,0.32 mg/L) regeneration culture medium on (regeneration culture medium is in MS substratum, add 30 g/L sucrose, 4.5 g/L agar, 0.3 mg/L BA and 0.01 mg/L NAA).The situation of test adventitious bud inducing regeneration.Above-mentioned containing different concns DA-6(0,0.01,0.08,0.16,0.32 mg/L) regeneration culture medium substratum collocation method with embodiment 1.The culture condition of adventitious bud inducing regeneration is be seeded in by dissimilar echinacea purpurea explant in regeneration culture medium suitable separately, it is 25 ± 4 DEG C in culture temperature, intensity of illumination is 1000 ~ 2500 lx, and light application time is cultivate 35d under the condition of 10 ~ 16 h/d.
The results are shown in Table 2 and Fig. 2.As can be seen from table 2 and Fig. 2: obviously DA-6 can stimulate leaf explant to regenerate, and optimum concn is 0.16 mg/L.Compare blade, the concentration that petiole needs is lower, is 0.08 mg/L.When DA-6 concentration is 0.01 mg/L, the regeneration effect of root explant improves slightly, but, when DA-6 concentration seriously suppresses regeneration higher than just starting during 0.01 mg/L.
Table 2 different concns DA-6 is on the impact of triploid explant regeneration indefinite bud
Note: the numerical value of each row different letter representation below significant difference (P<0.05) in Duncan test in table.
Embodiment 3
S1. the preparation of explant: with embodiment 1.
S2 tetraploid echinacea purpurea blade, petiole and root explant are inoculated into respectively containing different concns DA-6(0,0.01,0.08,0.16,0.32 mg/L) regeneration culture medium on (regeneration culture medium is in MS substratum, add 30 g/L sucrose, 4.5 g/L agar, 0.3 mg/L BA and 0.01 mg/L NAA).The situation of test adventitious bud inducing regeneration.Above-mentioned containing different concns DA-6(0,0.01,0.08,0.16,0.32 mg/L) regeneration culture medium substratum collocation method with embodiment 1.The culture condition of adventitious bud inducing regeneration is be seeded in by dissimilar echinacea purpurea explant in regeneration culture medium suitable separately, it is 25 ± 4 DEG C in culture temperature, intensity of illumination is 1000 ~ 2500 lx, and light application time is cultivate 35 d under the condition of 10 ~ 16 h/d.
The results are shown in Table 3 and Fig. 3.Leaf explant regenerated adventitious bud can be stimulated, optimum concn 0.08 mg/L as can be seen from table 3 and Fig. 3: DA-6.Comparatively speaking, the DA-6 concentration that petiole and root explant regeneration reach required for best regeneration effect is lower, is 0.01 mg/L.DA-6 concentration high to 0.32 mg/L time, inhibit blade and root explant regeneration.As for petiole explant, 0.16 mg/L or greater concn cause negative impact.
Table 3 different concns DA-6 is on the impact of tetraploid explant regeneration indefinite bud
Note: the numerical value of each row different letter representation below significant difference (P<0.05) in Duncan test in table.
Embodiment 4
S1. the preparation of explant: with embodiment 1.
S2 the echinacea purpurea blade of hexaploid, petiole and root explant are inoculated into respectively containing different concns DA-6(0,0.01,0.08,0.16,0.32 mg/L) regeneration culture medium on (regeneration culture medium is in MS substratum, add 30g/L sucrose, 4.5g/L agar, 0.3 mg/L BA and 0.01 mg/L NAA).The situation of test adventitious bud inducing regeneration.Above-mentioned containing different concns DA-6(0,0.01,0.08,0.16,0.32 mg/L) regeneration culture medium substratum collocation method with embodiment 1.The culture condition of adventitious bud inducing regeneration is be seeded in by dissimilar echinacea purpurea explant in regeneration culture medium suitable separately, it is 25 ± 4 DEG C in culture temperature, intensity of illumination is 1000 ~ 2500 lx, and light application time is cultivate 35 d under the condition of 10 ~ 16 h/d.
The results are shown in Table 4 and Fig. 4.Result as can be seen from table 4 and Fig. 4: DA-6 can promote blade and petiole explant regeneration, and optimum concn is 0.08 mg/L and 0.16 mg/L respectively.On the contrary, add the regeneration that DA-6 does not promote root explant, and the DA-6 of concentration 0.08 mg/L or greater concn suppresses regeneration.In this experiment, we find, the performance of hexaploid explant is different with those explants tested before, after adding the DA-6 of higher concentration, create a large amount of callus, and the callus that the callus of root explant and petiole explant generation produces than blade is many.
Table 4 different concns DA-6 is on the impact of hexaploid explant regeneration indefinite bud
Note: the numerical value of each row different letter representation below significant difference (P<0.05) in Duncan test in table.
Discuss: well-known, in prior art, the plant-growth regulator being generally used for tissue culture evoking adventive bud is BA and NAA, the former is as cell fission usually irritation cell division, as the key factor of regeneration induction approach.In echinacea purpurea, BA is widely used in evoking adventive bud equally.Result display of the present invention, by adding DA-6 in regeneration culture medium, can make the effect of BA regeneration induction be greatly enhanced.As plant-growth regulator, the working concentration of DA-6 is low (0.01 mg/L), very large to the best use of concentration difference of different explants, Different Ploidy material.The explant that but obviously ploidy is high is more responsive to DA-6, this is greatly unexpected---because just do not find in the early time, the higher BA concentration of material require of Hyperploidy is carried out regeneration induction indefinite bud or is promoted lateral bud growth (R. Chen, X. L. Chen, Q. L. Li, Y. S. Yang and H. Wu, Micropropagation by repeatedly inducing axillary bud formation of different gene dosage purple coneflower plants. Proceedings of The 2012 International Conference on Biomedical Engineering and Biotechnology, 28-30 May, 2012, Macao, pp.1056-1059.).
So far the data applied in tissue culture of DA-6 is little.Result of study display of the present invention, DA-6 plays very positive effect on adventitious bud inducing.This new plant growth regulator be worthy to be popularized very much other plant tissue culturing system in attempt.
It should be noted that, except above embodiment, the present invention can also have other embodiments and distortion, and what more than exemplify is only specific embodiments of the invention.All distortion that all those of ordinary skill in the art can directly derive from content disclosed by the invention or associate, all should think protection scope of the present invention.
Claims (8)
1. an echinacea purpurea adventitious shoot regeneration substratum, it is characterized in that, on the basis of MS formula except adding 15 ~ 60 g/L sucrose, 3 ~ 9 g/L agar, 0.1 ~ 1.5 mg/L BA and 0.01 ~ 0.2 mg/L NAA, also with the addition of diethylamino ethanol caproate especially, i.e. DA-6;
The amount of the DA-6 added in described diploid or the regeneration culture medium needed for triploid echinacea purpurea leaf explant is 0.01 ~ 0.32 mg/L;
The amount of the DA-6 added in tetraploid or the regeneration culture medium needed for hexaploid echinacea purpurea leaf explant is 0.01 ~ 0.16 mg/L;
The amount of the DA-6 added in diploid or the regeneration culture medium needed for triploid echinacea purpurea petiole explant is 0.01 ~ 0.16 mg/L;
The amount of the DA-6 added in the regeneration culture medium needed for tetraploid echinacea purpurea petiole explant is 0.01 mg/L;
The amount of the DA-6 added in the regeneration culture medium needed for hexaploid echinacea purpurea petiole explant is 0.08 ~ 0.16 mg/L;
The amount of the DA-6 added in diploid or the regeneration culture medium needed for triploid echinacea purpurea root explant is 0.01 mg/L;
The amount of the DA-6 added in the regeneration culture medium needed for tetraploid echinacea purpurea root explant is 0.01 ~ 0.16 mg/L.
2. regeneration culture medium according to claim 1, is characterized in that, the amount of the DA-6 added in described diploid or the regeneration culture medium needed for triploid echinacea purpurea leaf explant is 0.08 ~ 0.16 mg/L; The amount of the DA-6 added in tetraploid or the regeneration culture medium needed for hexaploid echinacea purpurea leaf explant is 0.08 ~ 0.16 mg/L.
3. regeneration culture medium according to claim 2, is characterized in that, the amount of the DA-6 added in described diploid or the regeneration culture medium needed for triploid echinacea purpurea leaf explant is 0.16 mg/L; The amount of the DA-6 added in tetraploid or the regeneration culture medium needed for hexaploid echinacea purpurea leaf explant is 0.08 mg/L.
4. regeneration culture medium according to claim 1, is characterized in that, the amount of the DA-6 added in described diploid or the regeneration culture medium needed for triploid echinacea purpurea petiole explant is 0.01 ~ 0.08 mg/L; The amount of the DA-6 added in the regeneration culture medium needed for hexaploid echinacea purpurea petiole explant is 0.16 mg/L.
5. regeneration culture medium according to claim 4, it is characterized in that, the amount of the DA-6 added in the regeneration culture medium needed for described diploid echinacea purpurea petiole explant is 0.08 mg/L; The amount of the DA-6 added in the regeneration culture medium needed for described triploid echinacea purpurea petiole explant is 0.01 mg/L.
6. regeneration culture medium according to claim 1, it is characterized in that, the amount of the DA-6 added in the regeneration culture medium needed for described tetraploid echinacea purpurea root explant is 0.01 ~ 0.08 mg/L.
7. regeneration culture medium according to claim 6, it is characterized in that, the amount of the DA-6 added in the regeneration culture medium needed for described tetraploid echinacea purpurea root explant is 0.01 mg/L.
8. one kind is improved the cultural method of echinacea purpurea explant regeneration indefinite bud, it is characterized in that, the needs of the component of the regeneration culture medium needed for echinacea purpurea explant in Different Ploidy described in 1 and source are wanted according to right, dissimilar echinacea purpurea explant is seeded in regeneration culture medium suitable separately, it is 25 ± 4 DEG C in culture temperature, intensity of illumination is 1000 ~ 2500 lx, and light application time is cultivate 18 ~ 60 d under the condition of 10 ~ 16 h/d.
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| CN104365475B (en) * | 2014-11-13 | 2016-09-28 | 华南农业大学 | A kind of screening technique obtaining hexaploid Echinacea |
| CN104365481B (en) * | 2014-11-13 | 2016-03-30 | 华南农业大学 | A kind of method improving colchicine-induced triploid Echinacea chromosome doubling efficiency |
| CN104429950B (en) * | 2014-11-14 | 2016-08-24 | 佛山市顺德区今日景艺生物科技有限公司 | Phalaenopsis Blume culture medium and cultural method |
| CN105052745A (en) * | 2015-08-24 | 2015-11-18 | 华南农业大学 | Method for promoting growth of echinacea purpurea regeneration buds |
| CN105052746A (en) * | 2015-08-24 | 2015-11-18 | 华南农业大学 | Echinacea adventitious bud regeneration culture method |
| CN106258978A (en) * | 2016-08-30 | 2017-01-04 | 华南农业大学 | One step obtains culture medium and the tissue culture method thereof of complete Echinacea regrowth |
| CN115197960A (en) * | 2022-08-16 | 2022-10-18 | 华南农业大学 | Composition and method for establishing genetic transformation system by taking echinacea purpurea petiole as explant |
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