CN103430842B - A kind of Quick propagation method of hybrid orchid tissue culture - Google Patents
A kind of Quick propagation method of hybrid orchid tissue culture Download PDFInfo
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Abstract
A kind of method that the invention provides hybrid orchid tissue-culturing quick-propagation, after sequentially passing through the inducing culture of bud, the enrichment culture of Multiple Buds, the successive transfer culture of Multiple Buds and root culture, obtains whole plant, can bottle outlet seedling exercising, and transplant.The method processing ease of the present invention, production cost is low, free from environmental pollution, it is possible to achieve large-scale production.By the hybrid orchid seedling that the present invention cultivates, its stabilization characteristics of genetics, maintain the characteristic of parent, possess and include invariance, less investment, output is high, the cycle is short many advantages.
Description
Technical field
The invention belongs to tissue culture rapid propagation technique field, be specifically related to a kind of Quick propagation method of hybrid orchid tissue culture.
Background technology
Hybrid orchid is the novel Cymbidium ensifolium (L.) Sw. of class selected by hybrid cymbidium and state's orchid artificial hybridization.The character such as the flower size of hybrid orchid, plant height, plant type, leaf blade size, between hybrid cymbidium and state orchid, have significantly high ornamental value, wide market.From annual flower market in recent years, supply falls short of demand for hybrid orchid, and some fine work hybrid orchid prices are high, and what have even sells for several thousand yuan of every basin, but the supply of current hybrid orchid relies primarily on import.Therefore, carry out hybrid orchid tissue-culturing quick-propagation research, improve sapling multiplication speed, the popularization and application to hybrid orchid, and the demand meeting market and be of great practical significance.
Summary of the invention
In view of this, technical problem solved by the invention there are provided a kind of Quick propagation method of hybrid orchid tissue culture, carry out hybrid orchid tissue culture by the approach of Multiple Buds and be obtained in that the test tube Seedling of a large amount of stabilization characteristics of genetics, keep good parental identity, and possess and include invariance, less investment, output is high, the cycle is short many advantages.
The present invention solves above-mentioned technical problem by the following technical programs:
A kind of Quick propagation method of hybrid orchid tissue culture, comprises the following steps:
1) sterilization of outer implant: take the sprouting of raw robust growth, high 5～15cm then, sprouting is cut by the place of the most base portion of maternal plant, removes impurity and leaf that upper end has been launched, leave behind the sprout of total length 4～5cm, running water 20min, with ethanol and 0.1%HgCl on superclean bench2Solution carries out process and realizes once sterilizing, then uses aseptic water washing 5～8 times again, then carefully peels off the outer quilt encased with tweezers piecewise, until exposing lateral bud, the stem section having lateral bud being cut down, then using 0.1%HgCl2Solution sterilization 6～8min carries out secondary sterilization, then with aseptic water washing 5～8 times, aseptic filter paper blots residual moisture, and is inoculated on inducing culture；
2) induction of Multiple Buds: adopting inducing culture to cultivate 15～20 days, lateral bud turns green increase；Then through 30～40 days, sprout formation also grew；In incubation, intensity of illumination 2000～2500lx, illumination 10～12 hour/day, temperature 25～28 DEG C；
3) propagation of Multiple Buds: utilize the Multiple Buds that lateral bud culture obtains, transfers to adventitious buds proliferation culture medium after clip partial blade, cultivate 40～60 days, it is thus achieved that proliferation times is the propagation Multiple Buds of 4.0～6.0；Condition of culture is: intensity of illumination 2000～2500lx, illumination 10～12 hour/day, temperature 25～28 DEG C；
4) successive transfer culture: adopt subculture medium to carry out successive transfer culture, every 40～60 days subculture multiplication 1 time, and subculture number is controlled within 15 times；Condition of culture is: intensity of illumination 2000～2500lx, illumination 10～12 hour/day, temperature 25～28 DEG C；
5) root culture: after Regenerated plant reaches suitable quantity, adopts root media to carry out Rooting and hardening-off culture, cultivates and obtain Seedling of taking root in 40～60 days.Condition of culture is: intensity of illumination 2000～2500lx, illumination 10～12 hour/day, temperature 25～28 DEG C；
6) test tube transplantation of seedlings: selecting annual 3～May is the season that bottle outlet is transplanted, or provide similar 3～May natural conditions growing environment carry out bottle outlet transplanting；Before transplanting, test tube Seedling is positioned over seedling exercising 10-20 days in the greenhouse of tool nature light scattering, then takes out seedling from test tube, cleans the culture medium of root, and be put in potassium permanganate solution and soak 3-5 minute, after taking-up with import water moss implantation in the plastic cup basin of bore 4-6cm；Keeping greenhouse ventilation, humidity is 70～80%, and temperature is maintained at more than 15 DEG C to room temperature range, and blower fan, cascade must be used to lower the temperature higher than 30 DEG C.
Preferably, described inducing culture based component is 1/2MS+6-BA (6-benzyl aminoadenine) 1.0～3.0/L+NAA (naphthalene acetic acid) 0.05～0.10/L+10.0～20.0% Sucus Cocois+3.0～5.0g/L activated carbon；The sugary 30g/L of culture medium, agar 0.7%, pH value 5.5-5.8.
Preferably, described adventitious buds proliferation culture medium is 1/2MS+6-BA (6-benzyl aminoadenine) 1.0～3.0/L+AD (adenine) 1.0～3.0/L+10.0～20.0% Sucus Cocois+3.0～5.0g/L activated carbon.
Preferably, described subculture medium is 1/2MS+6-BA (6-benzyl aminoadenine) 1.0～3.0/L+AD (adenine) 1.0～3.0/L+10.0～20.0% Sucus Cocois+3.0～5.0g/L activated carbon.
Preferably, containing 1/2MS+NAA (naphthalene acetic acid) 0.5～1.0/L+10.0～20.0% Sucus Cocois+3.0～5.0g/L activated carbon in described root culture based component.
Preferably, in the sterilisation step of described outer implant, with ethanol and 0.1%HgCl on superclean bench2Solution carries out process and realizes once sterilizing and refer to and first use alcohol washes surface on superclean bench, and soaks 15-45s in the ethanol of 75%, uses 0.1%HgCl2Solution sterilization 8～10min.
Preferably, in described test tube transplantation of seedlings, for soaking the potassium permanganate solution that potassium permanganate solution is 0.1% of seedling root.
Compared to prior art, having the beneficial effects that of the Quick propagation method of hybrid orchid tissue culture of the present invention: the method processing ease, production cost is low, free from environmental pollution, it is possible to accomplish scale production.By the hybrid orchid seedling that the present invention cultivates, its stabilization characteristics of genetics, maintain the characteristic of parent, possess and include invariance, less investment, output is high, the cycle is short many advantages.
Detailed description of the invention
In view of this, a kind of Quick propagation method of hybrid orchid tissue culture provided in the specific embodiment of the invention, specifically include following steps:
1) sterilization of outer implant: take the sprouting of raw robust growth, high 5～15cm then, sprouting is cut by the place of the most base portion of maternal plant, removes impurity and leaf that upper end has been launched, leave behind the sprout of total length 4～5cm, running water 20min, with ethanol and 0.1%HgCl on superclean bench2Solution carries out process and realizes once sterilizing, then uses aseptic water washing 5～8 times again, then carefully peels off the outer quilt encased with tweezers piecewise, until exposing lateral bud, the stem section having lateral bud being cut down, then using 0.1%HgCl2Solution sterilization 6～8min carries out secondary sterilization, then with aseptic water washing 5～8 times, aseptic filter paper blots residual moisture, and is inoculated on inducing culture；
2) induction of Multiple Buds: adopting inducing culture to cultivate 15～20 days, lateral bud turns green increase；Then through 30～40 days, sprout formation also grew；In incubation, intensity of illumination 2000～2500lx, illumination 10～12 hour/day, temperature 25～28 DEG C；
3) propagation of Multiple Buds: utilize the Multiple Buds that lateral bud culture obtains, transfers to adventitious buds proliferation culture medium after clip partial blade, cultivate 40～60 days, it is thus achieved that proliferation times is the propagation Multiple Buds of 4.0～6.0；Condition of culture is: intensity of illumination 2000～2500lx, illumination 10～12 hour/day, temperature 25～28 DEG C；
4) successive transfer culture: adopt subculture medium to carry out successive transfer culture, every 40～60 days subculture multiplication 1 time, and subculture number is controlled within 15 times；Condition of culture is: intensity of illumination 2000～2500lx, illumination 10～12 hour/day, temperature 25～28 DEG C；
5) root culture: after Regenerated plant reaches suitable quantity, adopts root media to carry out Rooting and hardening-off culture, cultivates and obtain Seedling of taking root in 40～60 days.Condition of culture is: intensity of illumination 2000～2500lx, illumination 10～12 hour/day, temperature 25～28 DEG C；
6) test tube transplantation of seedlings: selecting annual 3～May is the season that bottle outlet is transplanted, or provide similar 3～May natural conditions growing environment carry out bottle outlet transplanting；Before transplanting, test tube Seedling is positioned over seedling exercising 10-20 days in the greenhouse of tool nature light scattering, then takes out seedling from test tube, cleans the culture medium of root, and be put in potassium permanganate solution and soak 3-5 minute, after taking-up with import water moss implantation in the plastic cup basin of bore 4-6cm；Keeping greenhouse ventilation, humidity is 70～80%, and temperature is maintained at more than 15 DEG C to room temperature range, and blower fan, cascade must be used to lower the temperature higher than 30 DEG C.
The composition of the processing mode, condition of culture, incubation time and the culture medium that relate in each step above-mentioned all can carry out suitable adjustment according to specific needs.
Wherein, respectively cultivating when composition is determined, the content of each component wherein used can be adjusted according to actual cultivation situation, and the medium component used in said method and the content range of each composition are as follows:
In the induction step of Multiple Buds, containing 1/2MS+6-BA (6-benzyl aminoadenine) 1.0～3.0/L+NAA (naphthalene acetic acid) 0.05～0.10/L+10.0～20.0% Sucus Cocois+3.0～5.0g/L activated carbon in inducing culture based component；The sugary 30g/L of culture medium, agar 0.7%, pH value 5.5-5.8.
In the amplification step of Multiple Buds, adventitious buds proliferation culture medium contains 1/2MS+6-BA (6-benzyl aminoadenine) 1.0～3.0/L+AD (adenine) 1.0～3.0/L+10.0～20.0% Sucus Cocois+3.0～5.0g/L activated carbon.
In subculture step, containing 1/2MS+6-BA (6-benzyl aminoadenine) 1.0～3.0/L+AD (adenine) 1.0～3.0/L+10.0～20.0% Sucus Cocois+3.0～5.0g/L activated carbon in successive transfer culture based component.
In root culture step, containing 1/2MS+NAA (naphthalene acetic acid) 0.5～1.0/L+10.0～20.0% Sucus Cocois+3.0～5.0g/L activated carbon in root culture based component.
Further, since incubation is by the impact of the many factors such as such as temperature, illumination, humidity, thus, in each step of the present invention, processing mode, condition of culture, incubation time all can carry out suitable adjustment according to specific needs.
Wherein, in the sterilisation step of outer implant, with ethanol and 0.1%HgCl on superclean bench2Solution carries out process and realizes once sterilizing and refer to and first use alcohol washes surface on superclean bench, and soaks 15-45s in the ethanol of 75%, uses 0.1%HgCl2Solution sterilization 8～10min.It addition, in described test tube transplantation of seedlings, for soaking the potassium permanganate solution that potassium permanganate solution is 0.1% of seedling root.
For making the present invention easier to understand, specific embodiments of the invention are further illustrated below.
Embodiment 1, hybrid orchid tissue-culturing quick-propagation one:
Selecting the hybrid orchid of kind one to carry out tissue-culturing quick-propagation in the present embodiment, this kind is be commonly called as the hybrid orchid of " Korea S Miss ".
Take the sprouting of raw robust growth, high 5-15cm then, sprouting is cut by the place of the most base portion of maternal plant, the leaf that removal impurity and upper end have been launched, leave behind the sprout of total length 4-5cm, running water 20min, superclean bench is first used alcohol washes surface, and in the ethanol of 75%, soaks 30s, use 0.1%HgCl2Solution sterilization 8min, aseptic water washing 5 times, then carefully peel off the outer quilt encased with tweezers piecewise, until exposing lateral bud, the stem section having lateral bud being cut down, then uses 0.1%HgCl2Solution sterilization 6min, aseptic water washing 5 times, aseptic filter paper blots residual moisture, and it is seeded on inducing culture 1/2MS+6-BA (6-benzyl aminoadenine) 1.0/L+NAA (naphthalene acetic acid) 0.05/L+10.0% Sucus Cocois+3.0g/L activated carbon, intensity of illumination 2000～2500lx, illumination 12 hour/day, temperature 25～28 DEG C.Cultivating 45～60 days, sprout formation also grows.Utilize the Multiple Buds that lateral bud culture obtains, adventitious buds proliferation culture medium 1/2MS+6-BA (6-benzyl aminoadenine) 1.0/L+AD (adenine) 1.0/L+10.0% Sucus Cocois+3.0g/L activated carbon is transferred to after clip partial blade, at intensity of illumination 2000～2500lx, illumination 12 hour/day, under 25～28 DEG C of conditions of temperature, cultivate 40～60 days, it is thus achieved that Multiple Buds, proliferation times is up to 4.0.Every 40～60 days subculture multiplication 1 time, general subculture number is less than 15 times, subculture medium is 1/2MS+6-BA (6-benzyl aminoadenine) 1.0/L+AD (adenine) 1.0/L+10.0% Sucus Cocois+3.0g/L activated carbon, at intensity of illumination 2000～2500lx, illumination 12 hour/day, cultivates under 25～28 DEG C of conditions of temperature.After Regenerated plant reaches some, Rooting and hardening-off culture can be carried out, Rooting and hardening-off culture base is 1/2MS+NAA (naphthalene acetic acid) 0.5/L+10.0% Sucus Cocois+3.0g/L activated carbon, at intensity of illumination 2000～2500lx, illumination 12 hour/day, cultivate when temperature 25～28 DEG C, cultivate 40～60 days, obtain Seedling of taking root.Spring is the season that bottle outlet is transplanted, before transplanting, test tube Seedling is positioned over seedling exercising 15 days in the greenhouse of tool nature light scattering, then takes out seedling from test tube, clean the culture medium of root, and be put in the potassium permanganate solution of 0.1% and soak 5 minutes, after taking-up with import water moss implantation in the plastic cup basin of bore 4.8cm, and keep greenhouse ventilation, humidity is 70～80%, temperature is maintained at more than 15 DEG C, and blower fan, cascade must be used to lower the temperature higher than 30 DEG C, and transplanting survival rate is up to more than 90%.
Embodiment 2, hybrid orchid tissue-culturing quick-propagation two:
Selecting the hybrid orchid of kind two to carry out tissue-culturing quick-propagation in the present embodiment, this kind is be commonly called as the hybrid orchid of " Korea S beauty ".
Take the sprouting of raw robust growth, high 5-15cm then, sprouting is cut by the place of the most base portion of maternal plant, the leaf that removal impurity and upper end have been launched, leave behind the sprout of total length 4-5cm, running water 20min, superclean bench is first used alcohol washes surface, and in the ethanol of 75%, soaks 30s, use 0.1%HgCl2Solution sterilization 9min, aseptic water washing 6 times, then carefully peel off the outer quilt encased with tweezers piecewise, until exposing lateral bud, the stem section having lateral bud being cut down, then uses 0.1%HgCl2Solution sterilization 7min, aseptic water washing 6 times, aseptic filter paper blots residual moisture, and it is seeded on inducing culture 1/2MS+6-BA (6-benzyl aminoadenine) 2.0/L+NAA (naphthalene acetic acid) 0.075/L+15.0% Sucus Cocois+4.0g/L activated carbon, intensity of illumination 2000～2500lx, illumination 12 hour/day, temperature 25～28 DEG C.Cultivating 45～60 days, sprout formation also grows.Utilize the Multiple Buds that lateral bud culture obtains, adventitious buds proliferation culture medium 1/2MS+6-BA (6-benzyl aminoadenine) 2.0/L+AD (adenine) 2.0/L+15.0% Sucus Cocois+4.0g/L activated carbon is transferred to after clip partial blade, at intensity of illumination 2000～2500lx, illumination 12 hour/day, under 25～28 DEG C of conditions of temperature, cultivate 40～60 days, it is thus achieved that Multiple Buds, proliferation times is up to 5.0.Every 40～60 days subculture multiplication 1 time, general subculture number is less than 15 times, subculture medium is 1/2MS+6-BA (6-benzyl aminoadenine) 2.0/L+AD (adenine) 2.0/L+15.0% Sucus Cocois+4.0g/L activated carbon, at intensity of illumination 2000～2500lx, illumination 12 hour/day, cultivates under 25～28 DEG C of conditions of temperature.After Regenerated plant reaches some, Rooting and hardening-off culture can be carried out, Rooting and hardening-off culture base is 1/2MS+NAA (naphthalene acetic acid) 0.75/L+15.0% Sucus Cocois+4.0g/L activated carbon, at intensity of illumination 2000～2500lx, illumination 12 hour/day, cultivate when temperature 25～28 DEG C, cultivate 40～60 days, obtain Seedling of taking root.Spring is the season that bottle outlet is transplanted, before transplanting, test tube Seedling is positioned over seedling exercising 15 days in the greenhouse of tool nature light scattering, then takes out seedling from test tube, clean the culture medium of root, and be put in the potassium permanganate solution of 0.1% and soak 5 minutes, after taking-up with import water moss implantation in the plastic cup basin of bore 4.8cm, and keep greenhouse ventilation, humidity is 70～80%, temperature is maintained at more than 15 DEG C, and blower fan, cascade must be used to lower the temperature higher than 30 DEG C, and transplanting survival rate is up to more than 90%.
Embodiment 3, hybrid orchid tissue-culturing quick-propagation three:
Selecting the hybrid orchid of kind three to carry out tissue-culturing quick-propagation in the present embodiment, this kind is be commonly called as the hybrid orchid of " Dongfanghong ".
Take the sprouting of raw robust growth, high 5-15cm then, sprouting is cut by the place of the most base portion of maternal plant, the leaf that removal impurity and upper end have been launched, leave behind the sprout of total length 4-5cm, running water 20min, superclean bench is first used alcohol washes surface, and in the ethanol of 75%, soaks 30s, use 0.1%HgCl2Solution sterilization 10min, aseptic water washing 8 times, then carefully peel off the outer quilt encased with tweezers piecewise, until exposing lateral bud, the stem section having lateral bud being cut down, then uses 0.1%HgCl2Solution sterilization 8min, aseptic water washing 8 times, aseptic filter paper blots residual moisture, and it is seeded on inducing culture 1/2MS+6-BA (6-benzyl aminoadenine) 3.0/L+NAA (naphthalene acetic acid) 0.1/L+20.0% Sucus Cocois+5.0g/L activated carbon, intensity of illumination 2000～2500lx, illumination 12 hour/day, temperature 25～28 DEG C.Cultivating 45～60 days, sprout formation also grows.Utilize the Multiple Buds that lateral bud culture obtains, adventitious buds proliferation culture medium 1/2MS+6-BA (6-benzyl aminoadenine) 3.0/L+AD (adenine) 3.0/L+20.0% Sucus Cocois+5.0g/L activated carbon is transferred to after clip partial blade, at intensity of illumination 2000～2500lx, illumination 12 hour/day, under 25～28 DEG C of conditions of temperature, cultivate 40～60 days, it is thus achieved that Multiple Buds, proliferation times is up to 6.0.Every 40～60 days subculture multiplication 1 time, general subculture number is less than 15 times, subculture medium is 1/2MS+6-BA (6-benzyl aminoadenine) 3.0/L+AD (adenine) 3.0/L+20.0% Sucus Cocois+5.0g/L activated carbon, at intensity of illumination 2000～2500lx, illumination 12 hour/day, cultivates under 25～28 DEG C of conditions of temperature.After Regenerated plant reaches some, Rooting and hardening-off culture can be carried out, Rooting and hardening-off culture base is 1/2MS+NAA (naphthalene acetic acid) 0.1/L+20.0% Sucus Cocois+5.0g/L activated carbon, at intensity of illumination 2000～2500lx, illumination 12 hour/day, cultivate when temperature 25～28 DEG C, cultivate 40～60 days, obtain Seedling of taking root.Spring is the season that bottle outlet is transplanted, before transplanting, test tube Seedling is positioned over seedling exercising 15 days in the greenhouse of tool nature light scattering, then takes out seedling from test tube, clean the culture medium of root, and be put in the potassium permanganate solution of 0.1% and soak 5 minutes, after taking-up with import water moss implantation in the plastic cup basin of bore 4.8cm, and keep greenhouse ventilation, humidity is 70～80%, temperature is maintained at more than 15 DEG C, and blower fan, cascade must be used to lower the temperature higher than 30 DEG C, and transplanting survival rate is up to more than 90%.
Compared to prior art, the Quick propagation method of hybrid orchid tissue culture disclosed in aforesaid way, processing ease, production cost is low, free from environmental pollution, it is possible to accomplish scale production.By the hybrid orchid seedling that the present invention cultivates, its stabilization characteristics of genetics, maintain the characteristic of parent, possess and include invariance, less investment, output is high, the cycle is short many advantages.
Finally be should be noted that; above example is only in order to illustrate technical scheme but not limiting the scope of the invention; although the present invention being explained in detail with reference to preferred embodiment; it will be understood by those within the art that; technical scheme can be modified or equivalent replacement, without deviating from the spirit and scope of technical solution of the present invention.
Claims (3)
1. a Quick propagation method of hybrid orchid tissue culture, it is characterised in that the method comprises the following steps:
1) sterilization of outer implant: take the sprouting of raw robust growth, high 5～15cm then, sprouting is cut by the place of the most base portion of maternal plant, removes impurity and leaf that upper end has been launched, leave behind the sprout of total length 4～5cm, running water 20min, with ethanol and 0.1%HgCl on superclean bench2Solution carries out process and realizes once sterilizing, then uses aseptic water washing 5～8 times again, then carefully peels off the outer quilt encased with tweezers piecewise, until exposing lateral bud, the stem section having lateral bud being cut down, then using 0.1%HgCl2Solution sterilization 6～8min carries out secondary sterilization, then with aseptic water washing 5～8 times, aseptic filter paper blots residual moisture, and is inoculated on inducing culture；
2) induction of Multiple Buds: adopting inducing culture to cultivate 15～20 days, lateral bud turns green increase；Then through 30～40 days, sprout formation also grew；In incubation, intensity of illumination 2000～2500lx, illumination 10～12 hour/day, temperature 25～28 DEG C；Containing 1/2MS+6-BA (6-benzyl aminoadenine) 1.0～3.0/L+NAA (naphthalene acetic acid) 0.05～0.10/L+10.0～20.0% Sucus Cocois+3.0～5.0g/L activated carbon in described inducing culture based component；The sugary 30g/L of culture medium, agar 0.7%, pH value 5.5-5.8;
3) propagation of Multiple Buds: utilize the Multiple Buds that lateral bud culture obtains, transfers to adventitious buds proliferation culture medium after clip partial blade, cultivate 40～60 days, it is thus achieved that proliferation times is the propagation Multiple Buds of 4.0～6.0；Condition of culture is: intensity of illumination 2000～2500lx, illumination 10～12 hour/day, temperature 25～28 DEG C；Described adventitious buds proliferation culture medium contains 1/2MS+6-BA (6-benzyl aminoadenine) 1.0～3.0/L+AD (adenine) 1.0～3.0/L+10.0～20.0% Sucus Cocois+3.0～5.0g/L activated carbon;
4) successive transfer culture: adopt subculture medium to carry out successive transfer culture, every 40～60 days subculture multiplication 1 time, and subculture number is controlled within 15 times；Condition of culture is: intensity of illumination 2000～2500lx, illumination 10～12 hour/day, temperature 25～28 DEG C；Containing 1/2MS+6-BA (6-benzyl aminoadenine) 1.0～3.0/L+AD (adenine) 1.0～3.0/L+10.0～20.0% Sucus Cocois+3.0～5.0g/L activated carbon in described successive transfer culture based component;
5) root culture: after Regenerated plant reaches suitable quantity, adopts root media to carry out Rooting and hardening-off culture, cultivates and obtain Seedling of taking root in 40～60 days；Condition of culture is: intensity of illumination 2000～2500lx, illumination 10～12 hour/day, temperature 25～28 DEG C；Containing 1/2MS+NAA (naphthalene acetic acid) 0.5～1.0/L+10.0～20.0% Sucus Cocois+3.0～5.0g/L activated carbon in described root culture based component;
6) test tube transplantation of seedlings: selecting annual 3～May is the season that bottle outlet is transplanted, or provide similar 3～May natural conditions growing environment carry out bottle outlet transplanting；Before transplanting, test tube Seedling is positioned over seedling exercising 10-20 days in the greenhouse of tool nature light scattering, then takes out seedling from test tube, cleans the culture medium of root, and be put in potassium permanganate solution and soak 3-5 minute, after taking-up with import water moss implantation in the plastic cup basin of bore 4-6cm；Keeping greenhouse ventilation, humidity is 70～80%, and temperature is maintained at more than 15 DEG C to room temperature range, and blower fan, cascade must be used to lower the temperature higher than 30 DEG C.
2. Quick propagation method of hybrid orchid tissue culture as claimed in claim 1, it is characterised in that: in the sterilisation step of described outer implant, with ethanol and 0.1%HgCl on superclean bench2Solution carries out process and realizes once sterilizing and refer to and first use alcohol washes surface on superclean bench, and soaks 15-45s in the ethanol of 75%, uses 0.1%HgCl2Solution sterilization 8～10min.
3. Quick propagation method of hybrid orchid tissue culture as claimed in claim 1, it is characterised in that: in described test tube transplantation of seedlings, for soaking the potassium permanganate solution that potassium permanganate solution is 0.1% of seedling root.
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| CN111758561B (en) * | 2020-07-24 | 2022-04-15 | 广东省农业科学院环境园艺研究所 | Sterile sowing and breeding method of hybrid orchid seeds |
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