CN110178733B - Seedling rapid propagation culture medium and seedling rapid propagation method for cymbidium sinense protocorm - Google Patents
Seedling rapid propagation culture medium and seedling rapid propagation method for cymbidium sinense protocorm Download PDFInfo
- Publication number
- CN110178733B CN110178733B CN201910641383.3A CN201910641383A CN110178733B CN 110178733 B CN110178733 B CN 110178733B CN 201910641383 A CN201910641383 A CN 201910641383A CN 110178733 B CN110178733 B CN 110178733B
- Authority
- CN
- China
- Prior art keywords
- culture
- protocorm
- culture medium
- differentiation
- iaa
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Developmental Biology & Embryology (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention relates to the technical field of tissue culture, in particular to a seedling rapid propagation culture medium and a seedling rapid propagation method for cymbidium sinense protocorms. The seedling rapid propagation culture medium comprises an elongation propagation culture medium: MS culture medium containing sucrose, agar powder, NAA, and ZT; differentiation medium: MS culture medium containing sucrose, agar powder, IAA, and 6-BA; rooting culture medium: MS culture medium containing sucrose, agar powder, IAA, and 6-BA. Under the condition of tissue culture, the protocorm is regulated and controlled by hormone, the growth speed of the protocorm is high, the elongation and proliferation coefficient of the branch shape reaches 8.67, the protocorm is differentiated to generate a corm coefficient which reaches 5.14, namely 1 protocorm can generate about 45 healthy seedlings with complete root, stem and leaf after subculture and differentiation culture.
Description
Technical Field
The invention relates to the technical field of tissue culture, in particular to a seedling rapid propagation culture medium and a seedling rapid propagation method for cymbidium sinense protocorms.
Background
Mountain orchid Oreorchis patents (Lindl.) Lindl is perennial yin-growing herbaceous plant of mountain orchid family, and is widely distributed in southwest, China, northwest and northeast of China, and has little distribution from Korean peninsula to Siberian area, Japan and the like. Its dry bulb is sweet, pungent, cold and slightly toxic. Has the effects of nourishing yin, clearing away the lung-heat, eliminating phlegm and relieving cough, is used for carbuncle, infantile malnutrition, scrofula and innominate toxic swelling, but the medicinal value of the sansevieria trifasciata is not regarded as important for a long time, and the sansevieria trifasciata is treated as a pseudobulbus cremastrae seu pleiones or is used as a conventional product for being used as a medicine. Along with the increase of the demand of the sanlan in recent years and the rise of the price, the wild resources of the sanlan are destructively damaged, the sanlan is listed as a tertiary protection plant of Jilin province, and related researches on resource development, pharmacodynamic components, pharmacological activity, wild domestication, mycorrhizal fungi and the like are more, but the key technical research reports of seedling breeding for restricting large-area artificial cultivation of the sanlan are few.
The tissue culture technique is a technique in which living isolated organs (such as roots, stems, leaves, stem segments, protoplasts), tissues or cells are placed in a culture medium under artificially created aseptic conditions and placed in an appropriate environment for continuous culture to obtain cells, tissues or individuals. The technology is widely applied to agriculture and biological and medical research.
In the tissue culture technique of cymbidium sinense, there is a published technique of propagation culture of protocorm by using pseudobulb or leaf as explant, for example, the Chinese patent with publication number CN107736245A discloses a rapid asexual propagation technique of cymbidium sinense, which uses pseudobulb or leaf or inflorescence stalk as explant to mass-produce cymbidium sinense test-tube plantlet. The proliferation coefficient is long when the proliferation is low, while the proliferation coefficient is high when the proliferation is carried out in the form of protocorm, but the proliferation is carried out in the form of protocorm by one more step of differentiating protocorm into corm.
Disclosure of Invention
In view of the above, the present invention provides a seedling rapid propagation medium and a seedling rapid propagation method for cymbidium sinense protocorm. The invention takes the protocorm generated by the 1-year-old protocorm as an explant, and respectively studies the influence of 4 kinds of auxin and mitogen on the propagation and differentiation of the protocorm and the influence of hormone combination on the dendritic elongation propagation and differentiation of the protocorm and the generation of the protocorm and the rooting of the protocorm, thereby forming a complete Shanlan protocorm approach seedling rapid propagation technical system.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention also provides a seedling rapid propagation culture medium for the cymbidium sinense protocorm, which comprises the following components:
elongation multiplication medium: MS culture medium containing 28-32 g/L sucrose and 4.0-5.0 g/L, NAA 0.5.5-1.5 mg/L, ZT 0.5.5-1.5 mg/L agar powder;
differentiation medium: an MS culture medium containing 28-32 g/L of sucrose, 4.0-5.0 g/L, IAA 2.0.0-3.0 mg/L of agar powder and 2.0-3.0 mg/L of 6-BA;
rooting culture medium: an MS culture medium containing 28-32 g/L of sucrose, 4.0-5.0 g/L, IAA 2.0.0-3.0 mg/L of agar powder and 0.05-0.15 mg/L of 6-BA.
Preferably, the seedling rapid propagation medium comprises:
elongation multiplication medium: MS culture medium containing sucrose 30g/L and agar powder 4.5g/L, NAA 1.0.0 mg/L, ZT 1.0.0 mg/L;
differentiation medium: MS culture medium containing sucrose 30g/L, agar powder 4.5g/L, IAA 2.5.5 mg/L, and 6-BA2.5 mg/L;
rooting culture medium: MS culture medium containing 30g/L sucrose, 4.5g/L, IAA 2.5.5 mg/L agar powder and 0.1 mg/L6-BA.
The invention also provides a seedling rapid propagation method of the cymbidium kanran makino protocorm, which comprises the following steps:
performing elongation proliferation culture on the protocorm explant by adopting an elongation proliferation culture medium to obtain an elongation-proliferated protocorm; the elongation proliferation culture medium is an MS culture medium containing 28-32 g/L of sucrose and 4.0-5.0 g/L, NAA 0.5.5-1.5 mg/L, ZT 0.5.5-1.5 mg/L of agar powder;
carrying out differentiation culture on the extended and proliferated protocorm by adopting a differentiation culture medium to obtain a corm; the differentiation culture medium is an MS culture medium containing 28-32 g/L of sucrose, 4.0-5.0 g/L, IAA 2.0.0-3.0 mg/L of agar powder and 2.0-3.0 mg/L of 6-BA;
carrying out rooting culture on the corm by adopting a rooting culture medium; the rooting medium is an MS medium containing 28-32 g/L of sucrose, 4.0-5.0 g/L, IAA 2.0.0-3.0 mg/L of agar powder and 0.05-0.15 mg/L of 6-BA.
Preferably, the elongation growth medium is an MS medium containing 30g/L of sucrose and 4.5g/L, NAA 1.0.0 mg/L, ZT 1.0.0 mg/L of agar powder;
the differentiation culture medium is an MS culture medium containing 30g/L of sucrose, 4.5g/L, IAA 2.5.5 mg/L of agar powder and 2.5mg/L of 6-BA;
the rooting culture medium is an MS culture medium containing 30g/L of sucrose, 4.5g/L, IAA 2.5.5 mg/L of agar powder and 0.1mg/L of 6-BA.
Preferably, the size of the explant is (2-4) × (4-6) mm.
Preferably, the size of the explant is 3X 5 mm.
Preferably, the protocorm is a protocorm produced from a 1-year-old bulb.
Preferably, the conditions for the elongation growth culture are: the culture temperature is 22-24 ℃, the light-dark period is 13-15 h/9-11 h, the illumination intensity is 1400-1600 lx, and the culture time is 50-60 d.
Preferably, the conditions of the elongation propagation culture are: the culture temperature is 23 ℃, the light-dark period is 14h/10h, the illumination intensity is 1500lx, and the culture time is 55 d.
Preferably, the conditions for the differentiation culture are: the culture temperature is 22-24 ℃, the light-dark period is 13-15 h/9-11 h, the illumination intensity is 1400-1600 lx, and the culture time is 50-60 d.
Preferably, the conditions of the differentiation culture are: the culture temperature is 23 ℃, the light-dark period is 14h/10h, the illumination intensity is 1500lx, and the culture time is 55 d.
Preferably, the rooting culture conditions are as follows: the culture temperature is 22-24 ℃, the light-dark period is 13-15 h/9-11 h, the illumination intensity is 1400-1600 lx, and the culture time is 50-60 d.
Preferably, the rooting culture conditions are: the culture temperature is 23 ℃, the light-dark period is 14h/10h, the illumination intensity is 1500lx, and the culture time is 55 d.
The invention provides a seedling rapid propagation culture medium and a seedling rapid propagation method for a cymbidium sinense protocorm. The seedling rapid propagation culture medium comprises an elongation propagation culture medium: MS culture medium containing 28-32 g/L sucrose and 4.0-5.0 g/L, NAA 0.5.5-1.5 mg/L, ZT 0.5.5-1.5 mg/L agar powder; differentiation medium: an MS culture medium containing 28-32 g/L of sucrose, 4.0-5.0 g/L, IAA 2.0.0-3.0 mg/L of agar powder and 2.0-3.0 mg/L of 6-BA; rooting culture medium: an MS culture medium containing 28-32 g/L of sucrose, 4.0-5.0 g/L, IAA 2.0.0-3.0 mg/L of agar powder and 0.05-0.15 mg/L of 6-BA. The invention has the technical effects that:
in the experiment, the influence of 4 kinds of auxin and mitogen on the proliferation and differentiation of the protocorm and the influence of hormone combination on the dendritic elongation, proliferation and differentiation of the protocorm and the rooting of the protocorm are respectively researched by taking the protocorm generated by the 1-year-old protocorm as an explant and taking MS as a basic culture medium. The result shows that NAA and ZT are beneficial to protocorm branch elongation proliferation, and the protocorm branch elongation proliferation coefficient reaches 8.67 when NAA is 1.0mg/L and ZT is 1.0 mg/L; IAA and 6-BA are beneficial to the differentiation of the protocorm into the corm, and the coefficient of the protocorm differentiated into the corm reaches 5.14 when IAA is 2.5mg/L + 6-BA2.5mg/L; IAA and 6-BA are beneficial to the rooting of the bulb generated by the differentiation of the protocorm, when IAA is 2.5mg/L and 6-BA is 0.1mg/L, the number of the generated bulb reaches 6.26 per bulb, and the weight of the bulb reaches 9.90mg per bulb.
The experiment takes the protocorm generated by the 1-year-old protocorm as an explant, and respectively researches the influence of 4 kinds of auxin and mitogen on the propagation and differentiation of the protocorm and the influence of hormone combination on the dendritic elongation propagation and differentiation of the protocorm and the generation of the protocorm and the rooting of the protocorm, so as to form a complete Shanlan protocorm approach seedling rapid propagation technical system.
Under natural conditions, the cymbidium goeringii protocorm is slow in branch-shaped proliferation growth and only differentiates 1 corm, and the natural propagation efficiency is low; under the condition of tissue culture, the protocorm is regulated and controlled by hormone, the growth speed of the protocorm is high, the elongation and proliferation coefficient of the branch shape reaches 8.67, the protocorm is differentiated to generate a corm coefficient which reaches 5.14, namely about 45 robust seedlings with complete roots, stems and leaves can be generated after 1 protocorm is subjected to subculture proliferation and differentiation culture.
Drawings
FIG. 1 shows dendritic elongation propagation of protocorms;
FIG. 2 shows protocorm differentiation to give corms;
FIG. 3 shows the differentiation of the bulb to give roots.
Detailed Description
The invention discloses a seedling rapid propagation culture medium and a seedling rapid propagation method for cymbidium sinense protocorms. It is expressly intended that all such similar substitutes and modifications which would be obvious to one skilled in the art are deemed to be included in the invention. While the methods and applications of this invention have been described in terms of preferred embodiments, it will be apparent to those of ordinary skill in the art that variations and modifications in the methods and applications described herein, as well as other suitable variations and combinations, may be made to implement and use the techniques of this invention without departing from the spirit and scope of the invention.
The seedling rapid propagation culture medium of the cymbidium arundinaceum protocorm and the seeds, culture medium components and reagents used in the seedling rapid propagation method can be purchased from the market.
The invention is further illustrated by the following examples:
example 1
Test materials and methods
1.1 test materials
The protocorm generated by 1-year-old corm is taken as an explant, and related documents are referred to in a corm disinfection and culture method (Wangping, Wangyujiao, Chenxuhui, and the like. research on the isolated culture conditions of the mycorrhizal fungi of the mountain orchid [ J ]. northern horticulture, 2012, (09):66-69.), and the explant is the protocorm with a growing point and is about 3 × 5mm in size.
1.2 Effect of auxin on the proliferative differentiation of protocorms
Taking MS +6-BA0.1mg/L + sucrose 30g/L + agar powder 4.5g/L as a basic culture medium, and respectively adding 2,4-D, NAA, IBA and IAA at 0.1mg/L, 2.0mg/L, 4.0mg/L, 6.0mg/L, 8.0mg/L and 10.0 mg/L; the culture temperature is 23 ℃, the light-dark period is 14h/10h, the illumination intensity is 1500lx, the culture time is 55d, each treatment is repeated for 3 times, each time 15 explants are repeated, the statistical indexes comprise the elongation proliferation and differentiation coefficient of protocorm branches, the protocorm differentiation generation bulb coefficient and the average rootage number of protocorm differentiation generation bulbs, the dendritic elongation proliferation and differentiation coefficient of protocorms is the dendritic elongation branch number of protocorms/the number of inoculated protocorms, the protocorm differentiation generation bulb coefficient is the corm number generated by elongation differentiation of protocorms branches/the dendritic elongation branch number of protocorms, and the average rootage number of protocorms generated by protocorm differentiation generation bulbs is the rootage number of protocorms/the bulb generated by protocorm differentiation.
1.3 Effect of mitogens on the proliferative differentiation of protocorms
Taking MS + NAA0.1mg/L + sucrose 30g/L + agar powder 4.5g/L as a basic culture medium, and respectively adding TDZ, ZT, 6-BA and KT at 0.1mg/L, 2.0mg/L, 4.0mg/L, 6.0mg/L, 8.0mg/L and 10.0 mg/L; the culture temperature is 23 ℃, the light-dark period is 14h/10h, the illumination intensity is 1500lx, the culture time is 55d, each treatment is repeated for 3 times, each time is 15 explants, and the statistical index is 1.2.
1.4 Effect of NAA and ZT combination on dendritic, elongating, proliferating and differentiating protocorm
By taking MS, 30g/L of cane sugar and 4.5g/L of agar powder as a basic culture medium, combining NAA and ZT which have good effects on elongation, proliferation and differentiation of protocorm branches, wherein the concentration of the NAA is 0.1mg/L, 1.0mg/L, 2.0mg/L, 3.0mg/L and 4.0mg/L, and the concentration of the ZT is 0.1mg/L, 1.0mg/L, 2.0mg/L, 3.0mg/L and 4.0 mg/L; the culture temperature is 23 ℃, the light-dark period is 14h/10h, the illumination intensity is 1500lx, the culture time is 55d, each treatment is repeated for 3 times, each time the explants are repeated for 15 times, and the statistical index is the elongation proliferation differentiation coefficient of the protocorm branch.
1.5 Effect of IAA and 6-BA combinations on protocorm differentiation to generate corms and corm rooting
MS, 30g/L of sucrose and 4.5g/L of agar powder are used as a basic culture medium, and IAA and 6-BA with good effect of generating corms and corm rooting by differentiating protocorms are combined, wherein the IAA concentration is 0.1mg/L, 2.5mg/L, 5.0mg/L, 7.5mg/L and 10.0mg/L, and the 6-BA concentration is 0.1mg/L, 2.5mg/L, 5.0mg/L, 7.5mg/L and 10.0 mg/L; the culture temperature is 23 ℃, the light-dark period is 14h/10h, the illumination intensity is 1500lx, the culture time is 55d, each treatment is repeated for 3 times, each time 15 explants are repeated, and the statistical indexes comprise the protocorm differentiation generation corm coefficient, the average rooting number and the average root weight of protocorms, wherein the average root weight is the total root weight/the total root weight of the corms.
1.6 data analysis
Experimental data was analyzed for variance using SAS 8.01.
2 results and analysis
2.1 Effect of auxin on the proliferative differentiation of protocorms
2.1.1 Effect of auxin on dendritic elongation and proliferation and differentiation of protocorm As shown in Table 1, the auxin significant in dendritic elongation and proliferation effect of protocorm was NAA, and reached a maximum value proliferation system of 4.15 at 2.0mg/L of NAA.
TABLE 1 Effect of auxin on dendritic elongation of protocorm
2.1.2 effects of auxin on the differentiation of protocorms into bulbs As shown in Table 2, the auxin with better effect on the differentiation of protocorms into bulbs was IAA, and the maximum differentiation coefficient of 0.75 was reached at IAA of 6.0 mg/L.
TABLE 2 Effect of auxin on protocorm differentiation to generate corms
2.1.3 Effect of auxin on the rooting of bulbs produced by protocorm differentiation As shown in Table 3, the auxin having a good effect on the rooting of bulbs produced by protocorm differentiation was IAA, and the maximum rooting number of 1.67 pieces/bulb was reached at IAA of 6.0 mg/L.
TABLE 3 Effect of auxin on the rooting of bulbs resulting from differentiation of protocorms
2.2 Effect of mitogens on the proliferative differentiation of protocorms
2.2.1 Effect of mitogen on dendritic elongation, proliferation and differentiation of protocorm As shown in Table 4, the mitogen which favors dendritic elongation and proliferation of protocorm is ZT, and the maximum proliferation coefficient of ZT 2.0mg/L is 2.08.
TABLE 4 Effect of mitogens on dendritic elongation, proliferation and differentiation of protocorms
2.2.2 effects of mitogen on the generation of bulbs by protocorm differentiation As shown in Table 5, the mitogen having a significant effect on the generation of bulbs by protocorm differentiation was 6-BA, which reached a maximum differentiation coefficient of 0.56 at 6-BA6.0 mg/L.
TABLE 5 Effect of mitogens on the differentiation of protocorms to give bulbs
2.2.3 Effect of mitogen on the rooting of bulbs upon the differentiation of protocorms As shown in Table 6, the mitogen having a good effect on the rooting of bulbs upon the differentiation of protocorms was 6-BA, which reached a maximum of 1.81 pieces/bulb at 6-BA6.0 mg/L.
TABLE 6 Effect of mitogens on the rooting of bulbs resulting from differentiation of protocorms
2.3 Effect of NAA and ZT combination on dendritic, elongating, proliferating and differentiating protocorm
The effect of the combination of NAA and ZT on dendritic elongation, proliferation and differentiation of the protocorm is shown in Table 7, when the NAA is 1.0mg/L, the dendritic elongation, proliferation and differentiation coefficient of the protocorm tends to increase and decrease along with the increase of the ZT concentration, when the NAA is 1.0mg/L and the ZT is 1.0mg/L, the dendritic elongation, proliferation effect of the protocorm is better, and as shown in FIG. 1, the proliferation coefficient reaches 8.67.
TABLE 7 Effect of NAA and ZT combinations on dendritic, elongational, proliferative and differentiative of protocorms
2.4 Effect of IAA and 6-BA combinations on protocorm differentiation to yield corms and corm rooting and root weight
2.4.1 influence of IAA and 6-BA combination on the generation of corms by protocorm differentiation is shown in Table 8, when IAA is 2.5mg/L, the coefficient of the protocorm generation corms by protocorm differentiation tends to increase and decrease with the increase of 6-BA concentration, when IAA is 2.5mg/L +6-BA2.5mg/L, the effect of the protocorms by differentiation into corms is better, and the differentiation coefficient reaches 5.14, as shown in FIG. 2.
TABLE 8 Effect of IAA and 6-BA combinations on protocorm differentiation to produce corms
2.4.2 the effect of IAA and 6-BA combinations on the rooting of bulbs from protocorm differentiation is shown in Table 9, with IAA of 2.5mg/L being beneficial to the rooting of bulbs from protocorm differentiation, the number of roots decreasing with increasing concentration of 6-BA, and with IAA of 2.5mg/L +6-BA of 0.1mg/L the number of roots of bulbs is the most, reaching 6.26 bulbs, as shown in FIG. 3.
TABLE 9 Effect of IAA and 6-BA combinations on protocorm differentiation to corm rooting (strips/corms)
2.4.3 influence of IAA and 6-BA combination on the bulb root weight produced by protocorm differentiation is shown in Table 10, IAA of 2.5mg/L is beneficial to root growth, the root weight is reduced along with the increase of 6-BA concentration, and the root weight produced by the bulb is maximum at IAA of 2.5mg/L +6-BA0.1mg/L and reaches 9.90 mg/strip.
TABLE 10 Effect of IAA and 6-BA combinations on bulb root weight resulting from protocorm differentiation (mg/stripe)
Discussion of 3
3.1 the cymbidium protocorm has high multiplication coefficient and is a relatively ideal multiplication material, but the protocorm directly obtained from soil is used as a primary inoculation material and has the problems of difficult disinfection and low survival rate, while the 1-year-old protocorm has smooth surface, easy disinfection and vigorous vitality, so that the 1-year-old protocorm is suitable as the primary inoculation material, and the sterile protocorm differentiated from the 1-year-old protocorm is used as a secondary multiplication material.
3.2 under the condition of tissue culture, the protocorm is regulated and controlled by hormone, the protocorm is proliferated in two modes of callus and branch elongation, the branch elongation proliferation mode is convenient for production operation and is a main proliferation mode of protocorm approach seedling rapid propagation.
3.3 the concentration of hormone which is the most effective in differentiating protocorms into bulbs and bulb rooting in the one-way test does not show the best differentiation under the combined conditions, but is the most effective when combined at lower concentrations, which may be the reason for the antagonistic effect at high hormone concentrations.
3.4 since the roots grow curly, the length is not easy to determine, but the diameter of the roots is approximately the same, so the rooting effect is measured by two indexes of the number of the roots and the average weight of the roots.
3.5 under natural conditions, the branch-shaped proliferation growth of the cymbidium sinense protocorm is slow, only 1 corm is differentiated, and the natural propagation efficiency is low; under the condition of tissue culture, the protocorm is regulated and controlled by hormone, the growth speed of the protocorm is high, the elongation and proliferation coefficient of the branch shape reaches 8.67, the protocorm is differentiated to generate a corm coefficient which reaches 5.14, namely about 45 robust seedlings with complete roots, stems and leaves can be generated after 1 protocorm is subjected to subculture proliferation and differentiation culture.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (10)
1. A seedling rapid propagation culture medium for cymbidium sinense protocorms is characterized by comprising the following culture media:
branched elongation and proliferation medium: 28-32 g/L of sucrose, 4.0-5.0 g/L of agar powder, 0.5-1.5 mg/L of NAA, 0.5-1.5 mg/L of ZT and MS culture medium;
differentiation medium: 28-32 g/L of sucrose, 4.0-5.0 g/L of agar powder, 2.0-3.0 mg/L of IAA and 2.0-3.0 mg/L of 6-BA and MS culture medium;
rooting culture medium: 28-32 g/L of sucrose, 4.0-5.0 g/L of agar powder, 2.0-3.0 mg/L of IAA and 0.05-0.15 mg/L of 6-BA plus MS culture medium.
2. A seedling rapid propagation medium according to claim 1, comprising:
branched elongation and proliferation medium: 30g/L of sucrose, 4.5g/L of agar powder, 1.0mg/L of NAA, 1.0mg/L of ZT and MS culture medium;
differentiation medium: 30g/L of sucrose, 4.5g/L of agar powder, 2.5mg/L of IAA, 2.5mg/L of 6-BA and MS culture medium;
rooting culture medium: 30g/L of sucrose, 4.5g/L of agar powder, 2.5mg/L of IAA, 0.1mg/L of 6-BA and MS culture medium.
3. A seedling rapid propagation method of a cymbidium protocorm is characterized by comprising the following steps:
performing branched elongation propagation culture on the protocorm explant by using a branched elongation propagation culture medium to obtain a branched elongation-propagated protocorm; the branched elongation proliferation culture medium comprises: 28-32 g/L of sucrose, 4.0-5.0 g/L of agar powder, 0.5-1.5 mg/L of NAA, 0.5-1.5 mg/L of ZT and MS culture medium;
carrying out differentiation culture on the protocorm after the branch-shaped elongation proliferation by adopting a differentiation culture medium to obtain a corm; the differentiation medium is as follows: 28-32 g/L of sucrose, 4.0-5.0 g/L of agar powder, 2.0-3.0 mg/L of IAA and 2.0-3.0 mg/L of 6-BA and MS culture medium;
carrying out rooting culture on the corm by adopting a rooting culture medium; the rooting culture medium comprises: 28-32 g/L of sucrose, 4.0-5.0 g/L of agar powder, 2.0-3.0 mg/L of IAA and 0.05-0.15 mg/L of 6-BA plus MS culture medium.
4. A seedling rapid propagation method according to claim 3, wherein the branched elongation propagation medium is: 30g/L of sucrose, 4.5g/L of agar powder, 1.0mg/L of NAA, 1.0mg/L of ZT and MS culture medium;
the differentiation medium is as follows: 30g/L of sucrose, 4.5g/L of agar powder, 2.5mg/L of IAA, 2.5mg/L of 6-BA and MS culture medium;
the rooting culture medium comprises: 30g/L of sucrose, 4.5g/L of agar powder, 2.5mg/L of IAA, 0.1mg/L of 6-BA and MS culture medium.
5. A rapid seedling propagation method according to claim 3, wherein the size of the explant is (2-4) × (4-6) mm.
6. A method for rapid propagation of a seedling as claimed in claim 3, wherein the protocorm is a 1 year old protocorm.
7. A method according to claim 3, wherein said branched elongation propagation culture is performed under conditions of: the culture temperature is 22-24 ℃, the light-dark period is 13-15 h/9-11 h, the illumination intensity is 1400-1600 lx, and the culture time is 50-60 d.
8. Method according to any of claims 3 to 7, wherein said branched elongation propagation culture is performed under conditions of: the culture temperature is 23 ℃, the light-dark period is 14h/10h, the illumination intensity is 1500lx, and the culture time is 55 d.
9. The method of claim 3, wherein the differentiation culture conditions are: the culture temperature is 22-24 ℃, the light-dark period is 13-15 h/9-11 h, the illumination intensity is 1400-1600 lx, and the culture time is 50-60 d.
10. The method of claim 3, wherein the rooting culture conditions are: the culture temperature is 22-24 ℃, the light-dark period is 13-15 h/9-11 h, the illumination intensity is 1400-1600 lx, and the culture time is 50-60 d.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910641383.3A CN110178733B (en) | 2019-07-16 | 2019-07-16 | Seedling rapid propagation culture medium and seedling rapid propagation method for cymbidium sinense protocorm |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910641383.3A CN110178733B (en) | 2019-07-16 | 2019-07-16 | Seedling rapid propagation culture medium and seedling rapid propagation method for cymbidium sinense protocorm |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110178733A CN110178733A (en) | 2019-08-30 |
| CN110178733B true CN110178733B (en) | 2021-02-09 |
Family
ID=67725865
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910641383.3A Active CN110178733B (en) | 2019-07-16 | 2019-07-16 | Seedling rapid propagation culture medium and seedling rapid propagation method for cymbidium sinense protocorm |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110178733B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112753575B (en) * | 2020-12-31 | 2023-01-17 | 湖北金水源农业开发有限公司 | High-yield Cremastra appendiculata seedling cultivation method |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100444722C (en) * | 2006-01-18 | 2008-12-24 | 贵州省农业科学院生物技术研究所 | Cyrtopterin tissue culturing method and fast reproduction thereof |
| CN105941145B (en) * | 2016-05-04 | 2018-05-15 | 中国农业科学院特产研究所 | A kind of germination method of mountain orchid species |
| CN109122325A (en) * | 2018-11-09 | 2019-01-04 | 翁源县天下泽雨农业科技有限公司 | A kind of aseptic seeding quick-breeding method of sword-leaved cymbidium seed |
-
2019
- 2019-07-16 CN CN201910641383.3A patent/CN110178733B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN110178733A (en) | 2019-08-30 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101213941B (en) | Fast replication and in-vitro conservation method for dendrobium | |
| Ray et al. | In vitro regeneration of brinjal (Solanum melongena L.) | |
| CN102907318B (en) | A kind of method utilizing the fast numerous pseudo-ginseng regeneration plant of bioreactor culture somatic embryo | |
| CN115537346A (en) | Mucillus mucilaginosus for promoting growth and differentiation of sansevieria trifasciata and application thereof | |
| KR100883188B1 (en) | Induction Method of Protocorm like Body from Phalaenopsis Adult Plant | |
| CN110178733B (en) | Seedling rapid propagation culture medium and seedling rapid propagation method for cymbidium sinense protocorm | |
| CN112042541B (en) | Method for propagating taxillus through somatic embryogenesis | |
| Pathak et al. | In vitro propagation and mass scale multiplication of a critically endangered epiphytic orchid, Gastrochilus calceolaris (Buch.-Ham ex JE Sm.) D. Don. | |
| Danial et al. | Response of running shoot tips of strawberry (Fragaria x ananasa) for in vitro propagation in Kurdistan region of Iraq | |
| Govindaraju et al. | In vitro propagation of banana (Musa sp-Rasthali variety) from sword suckers for its commercial production. | |
| CN110178734B (en) | Directional culture medium for cymbidium goeringii germinated seeds and directional culture seedling method | |
| CN109601387B (en) | Tissue culture propagation method of osmunda vachellii with GGB route induced by juvenile sporocyst group | |
| CN114931079B (en) | Application of endophytic fungus P-B313 in improving low phosphorus stress resistance of dendrobium nobile | |
| CN110679481A (en) | Method for cultivating tetraploid polygonum capitatum | |
| CN114375834B (en) | Culture method for inducing differentiation of blumea balsamifera root cells to generate adventitious buds in one step | |
| CN113598054B (en) | Rapid seedling culture method for tissue culture of stem tips of pachyrhizua angulatus | |
| CN113068611B (en) | Method for rapidly propagating seedlings by utilizing leaves of tetrapanax tetraphyllus | |
| Chaitanya et al. | Review on Propagation Techniques of Jasmine (Jasminum sambac (L.)) | |
| CN111990255B (en) | Method for inducing and regenerating leaf callus of kudzu vine root tissue culture seedling | |
| Liu et al. | Effects of different factors on adventitious bud induction from stem explants of Ludisia discolor | |
| Huang et al. | Chinese gooseberry, kiwifruit (Actinidia spp.) | |
| CN108782244B (en) | Tissue culture method for longzhuguo | |
| Badal et al. | Clonal Propagation of Strawberry (Fragaria x ananassa Duch.) through In vitro Runner Tip Culture through Incorporation of Growth Hormones | |
| KR20080104632A (en) | Proliferation method of phalaenopsis protocorm like body | |
| CN114456951B (en) | Horizontalium fungus strain for promoting growth of ginseng, and method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |