CN103548694B - Tissue culture and rapid propagation method for dracaena cochinchinensis - Google Patents
Tissue culture and rapid propagation method for dracaena cochinchinensis Download PDFInfo
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Abstract
The invention provides a tissue culture and rapid propagation method for dracaena cochinchinensis. The tissue culture and rapid propagation method comprises the following steps: (1) taking a dracaena cochinchinensis seed as an explant and disinfecting; (2) putting the disinfected explant into an MS (Murashige and Skoog) culture medium to be induced and burgeoned to obtain a sterile test-tube plantlet; (3) putting the sterile test-tube plantlet into the MS culture medium to be subjected to test-tube plantlet rapid propagation and culture to obtain cluster buds; (4) putting the cluster buds into an MS strong seedling culture medium to carry out strong seedling culture to obtain a strong plant; (5) putting the strong plant into an MS rooting culture medium to be cultured to obtain a complete seedling with roots; and (6) taking the complete seedling with the roots and carrying out domestication; and transplanting the seedling into a sand bed to grow for one month and then transplanting the seedling to a large field. With the adoption of the culture method, the growth coefficient of the cluster buds of the obtained dracaena cochinchinensis reaches 10-15 times; the rooting rate of an obtained tissue culture seedling is more than 95% and the survival rate of a transplanted seedling bed is more than 90%, so that the large-scale seedling growth problem of the dracaena cochinchinensis is solved effectively.
Description
Technical field
The present invention relates to a kind of method for propagation, particularly a kind of quick breeding method for tissue culture of swordleaf dragon tree.
Background technology
Swordleaf dragon tree have another name called thousand wood, dragon's blood tree, toe dragon tree; it is vulnerable species; belong to Chinese Second Class Key Protected Plant; the international nature of 2003 Nian Bei is classified as red list with the natural resources protection federation species incorporated into as being subject to endanger; being decided to be the endangered species in the Red Data Book of Cape Verde, is the Original plant of current domestic generally acknowledged domestic Resina Draconis.
Dragon's blood is a kind of famous and precious traditional Chinese medicine, has more than 1500 in the use history of China.Record according to Compendium of Material Medica, Resina Draconis is warm in nature, flat, and taste is sweet, salty, nontoxic, enters blood system, and return lung, spleen, kidney three warp, have promoting blood circulation and removing blood stasis, swelling and pain relieving, astringing to arrest bleeding, the remarkable efficacy such as softening and resolving hard mass, regenerating tissue to heal wond, is described as panacea of invigorating blood circulation.Modern medicine study confirm, dragon's blood have improve body microcirculation, adjustment body metabolism, improve the effects such as body's immunity.
Although dragon's blood use is at home with a long history, be rely on Southeast Asia and African import before 1972, price is very expensive always.Mr. Cai Xitao in 1973 found large stretch of dragon tree that can produce dragon's blood on the lime mountain that Lian County in the Meng, Simao, Yunnan area is domestic, just the respite resource pressure of China's dragon's blood, thus the history of domestic Resina Draconis that has been born.But the growth of dragon tree slowly, within 1 year, trunk increasing is slightly less than 1 centimetre, and Resina Draconis is again the red liquid of the injured rear outflow of dragon tree, and output is very limited.And along with the transformation of medical idea, increasing crowd tends to use Chinese Traditional Medicine to prevent and cure diseases, and the consumption of natural resources of Chinese medicinal materials sharply rises, and increasing medicinal plant is tending towards rare, even in imminent danger.In recent years; market is to the continuous expansion of Resina Draconis demand; current Resina Draconis price has increased to 1000-1300 unit/kg; cause people to implement irrational predatoriness to swordleaf dragon tree wild resource to fell; swordleaf dragon tree wild resource is increasingly exhausted; extinction in imminent danger, is classified as rare endangered plants by a lot of country in the world.
In order to strengthen the protection of Resina Draconis Original plant, preventing the extinction in imminent danger of swordleaf dragon tree, realizing the sustainable use of resource, needing the cultivating and growing wideling popularize swordleaf dragon tree.On seedling breeding, swordleaf dragon tree is by seed, cuttage or tillering propagation, but the proliferative speed of this conventional method is low, and required time is long, and the seedling quantity of gained is very limited, production is difficult to realize commerial growing.And adopt biotechnology tissue culture technique, reproduction speed and the quality of Dracaena cochinchinensis seeding can be improved effectively rapidly, realize the factorial seedling growth of swordleaf dragon tree high quality seedling, with the needs in satisfied production.
Summary of the invention
The object of this invention is to provide a kind of quick breeding method for tissue culture of swordleaf dragon tree, it can Fast-propagation go out a large amount of be applicable to transplanting excellent Dracaena cochinchinensis seeding, meet need of production.
The present invention achieves the above object by the following technical programs: a kind of quick breeding method for tissue culture of swordleaf dragon tree, comprises the following steps:
(1) selection of explant and sterilization: get swordleaf dragon tree seed as explant, successively with 2% liquid detergent aqueous solution soaking 5min, wire tap water 15-30min, with the addition of 2-3 and drip 100 milliliter of 0.1% mercuric chloride sterilization 8-10min of Tween-20, aseptic water washing 3-5 time, finally remove surface moisture with sterilized filter paper, obtain explant, wherein sterile water is through autoclaved distilled water;
(2) seed germination obtains in vitro cuttings: be inoculated in MS medium by the explant that step (1) obtains, be 23-27 DEG C in cultivation temperature, intensity of illumination 1500lux, light application time is cultivate 30 days under the condition of 12-14 hour/day, in vitro cuttings is obtained after seed sprouting, wherein contain heteroauxin IAA, 30g/L sucrose of 6-benzyladenine 6-BA, 0.2mg/L and the agar of 3.4g/L of 1.0mg/L in MS medium, the pH value of medium is 5.8;
(3) test-tube plantlet Multiple Buds Fast-propagation is cultivated: the in vitro cuttings obtained in step (2) is placed in MS propagating culture medium, at cultivation temperature 23-27 DEG C, intensity of illumination 1500lux, light application time is cultivate under the condition of 8-10 hour/day to obtain test-tube plantlet Multiple Buds in 30 days, wherein contain kinetin KT, 30g/L sucrose of heteroauxin IAA, 0.1-2.0mg/L and the agar of 3.4g/L of 6-benzyladenine 6-BA, 0.1-0.5mg/L of 1.0-5.0mg/L in MS propagating culture medium, the pH value of medium is 5.8;
(4) Multiple Buds strong seedling culture: the test-tube plantlet Multiple Buds obtained in step (3) is placed in MS strong seedling culture base, at cultivation temperature 23-27 DEG C, intensity of illumination 1500lux, light application time is cultivate under the condition of 12-14 hour/day to obtain healthy and strong plant in 20 days, wherein contain heteroauxin IAA, 30g/L sucrose of 6-benzyladenine 6-BA, 0.5-1.5mg/L and the agar of 3.4g/L of 1.0-3.0mg/L in MS strong seedling culture base, the pH value of medium is 5.8;
(5) healthy and strong plant culture of rootage: the healthy and strong plant obtained in step (4) is placed in MS root media, at cultivation temperature 23-27 DEG C, intensity of illumination 1500lux, light application time is cultivate the whole plant obtaining for 35 days being with root under the condition of 12-14 hour/day, wherein contain root-inducing powder ABT, 30g/L sucrose of heteroauxin IAA, 0.1-0.5mg/L and the agar of 3.4g/L of 0.1-1.0mg/L in MS root media, the pH value of medium is 5.8;
(6) hardening and transplanting: after step (5) obtains the whole plant being with root, bottle cap is opened in the indoor being 25 DEG C in room temperature, a small amount of running water is added in bottle, hardening 2-4 days, after surface horny is formed, seedling is taken out, clean root medium, be transplanted in husky bed immediately, grow one month in husky bed after, transplant land for growing field crops.
Outstanding advantages of the present invention is:
(1) biotechnology is adopted to carry out tissue-culturing quick-propagation to swordleaf dragon tree; by the breeding of swordleaf dragon tree Multiple Buds; a large amount of swordleaf dragon tree seedling being applicable to cultivating and growing can be cultivated at short notice; significantly improve growth coefficient and the seedling quality of Dracaena cochinchinensis seeding; accomplish scale production, meet the needs on producing.
(2) the 6-benzyladenine 6-BA added in MS propagating culture medium, heteroauxin IAA and kinetin KT belong to chemosynthesis material, and price inexpensively, greatly can reduce the cost consumption of swordleaf dragon tree tissue cultures, reach the object reduced costs;
The kinetin adding 6-benzyladenine 6-BA and 0.1-2.0mg/L of 1.0-5.0mg/L in MS propagating culture medium can promote the differentiation of Multiple Buds; Adding concentration is the growth that the somatotropin heteroauxin IAA of 0.1-0.5mg/L can promote Multiple Buds;
In MS strong seedling culture base, add concentration is that the heteroauxin IAA of 6-benzyladenine 6-BA and 0.5-1.5mg/L of 1.0-3.0mg/L can promote the propagation again of Multiple Buds and the Zhan Ye that grows tall;
MS root media combinationally uses somatotropin IAA and 0.1-0.5mg/L root-inducing powder ABT that concentration is 0.1-1.0mg/L, and can obtain the whole plant of band root, these plant can directly transplant husky bed after hardening.
(3) the swordleaf dragon tree adventitious buds proliferation coefficient adopting cultural method of the present invention to obtain reaches 10-15 doubly, Multiple Buds is after rejuvenation, be inoculated on the root media containing root-inducing powder IAA, ABT, the plantlet in vitro rooting rate obtained, more than 95%, transplants seedbed survival rate more than 90%; Fast, convenient, carry out swordleaf dragon tree tissue cultures efficiently, realize the large-scale production that swordleaf dragon tree group cultivates seedling.
Embodiment
Below in conjunction with embodiment, technical scheme of the present invention is further illustrated.
Embodiment 1
An example of the quick breeding method for tissue culture of swordleaf dragon tree of the present invention, comprises the following steps:
(1) selection of explant and sterilization: get swordleaf dragon tree seed as explant, successively with 2% liquid detergent aqueous solution soaking 5min, wire tap water 15-30min, with the addition of 2-3 and drip 100 milliliter of 0.1% mercuric chloride sterilization 8-10min of Tween-20, aseptic water washing 3-5 time, finally remove surface moisture with sterilized filter paper, obtain explant, wherein sterile water is through autoclaved distilled water;
(2) seed germination obtains in vitro cuttings: be inoculated in MS medium by the explant that step (1) obtains, be 23-27 DEG C in cultivation temperature, intensity of illumination 1500lux, light application time is cultivate 30 days under the condition of 12-14 hour/day, in vitro cuttings is obtained after seed sprouting, wherein contain heteroauxin IAA, 30g/L sucrose of 6-benzyladenine 6-BA, 0.2mg/L and the agar of 3.4g/L of 1.0mg/L in MS medium, the pH value of medium is 5.8;
(3) test-tube plantlet Multiple Buds Fast-propagation is cultivated: the in vitro cuttings obtained in step (2) is placed in MS propagating culture medium, at cultivation temperature 23-27 DEG C, intensity of illumination 1500lux, light application time is cultivate under the condition of 8-10 hour/day to obtain test-tube plantlet Multiple Buds in 30 days, wherein contain kinetin KT, 30g/L sucrose of heteroauxin IAA, 2.0mg/L and the agar of 3.4g/L of 6-benzyladenine 6-BA, 0.5mg/L of 1.0mg/L in MS propagating culture medium, the pH value of medium is 5.8, and adventitious buds proliferation coefficient is 9.2;
(4) Multiple Buds strong seedling culture: the test-tube plantlet Multiple Buds obtained in step (3) is placed in MS strong seedling culture base, at cultivation temperature 23-27 DEG C, intensity of illumination 1500lux, light application time is cultivate under the condition of 12-14 hour/day to obtain healthy and strong plant in 20 days, wherein contain heteroauxin IAA, 30g/L sucrose of 6-benzyladenine 6-BA, 0.5-1.5mg/L and the agar of 3.4g/L of 1.0-3.0mg/L in MS strong seedling culture base, the pH value of medium is 5.8, and the coefficient that Multiple Buds is bred again is 2.8;
(5) healthy and strong plant culture of rootage: the healthy and strong plant obtained in step (4) is placed in MS root media, at cultivation temperature 23-27 DEG C, intensity of illumination 1500lux, light application time is cultivate the whole plant obtaining for 35 days being with root under the condition of 12-14 hour/day, wherein contain root-inducing powder ABT, 30g/L sucrose of heteroauxin IAA, 0.1-0.5mg/L and the agar of 3.4g/L of 0.1-1.0mg/L in MS root media, the pH value of medium is 5.8, and rooting rate is 90.6%;
(6) hardening and transplanting: after step (5) obtains the whole plant being with root, bottle cap is opened in the indoor being 25 DEG C in room temperature, a small amount of running water is added in bottle, hardening 2-4 days, after surface horny is formed, seedling is taken out, clean root medium, be transplanted in husky bed immediately, grow one month in husky bed after, transplant land for growing field crops.
Embodiment 2
Another example of the quick breeding method for tissue culture of swordleaf dragon tree of the present invention, comprises the following steps:
(1) selection of explant and sterilization: get swordleaf dragon tree seed as explant, successively with 2% liquid detergent aqueous solution soaking 5min, wire tap water 15-30min, with the addition of 2-3 and drip 100 milliliter of 0.1% mercuric chloride sterilization 8-10min of Tween-20, aseptic water washing 3-5 time, finally remove surface moisture with sterilized filter paper, obtain explant, wherein sterile water is through autoclaved distilled water;
(2) seed germination obtains in vitro cuttings: be inoculated in MS medium by the explant that step (1) obtains, be 23-27 DEG C in cultivation temperature, intensity of illumination 1500lux, light application time is cultivate 30 days under the condition of 12-14 hour/day, in vitro cuttings is obtained after seed sprouting, wherein contain heteroauxin IAA, 30g/L sucrose of 6-benzyladenine 6-BA, 0.2mg/L and the agar of 3.4g/L of 1.0mg/L in MS medium, the pH value of medium is 5.8;
(3) test-tube plantlet Multiple Buds Fast-propagation is cultivated: the in vitro cuttings obtained in step (2) is placed in MS propagating culture medium, at cultivation temperature 23-27 DEG C, intensity of illumination 1500lux, light application time is cultivate under the condition of 8-10 hour/day to obtain test-tube plantlet Multiple Buds in 30 days, wherein contain kinetin KT, 30g/L sucrose of heteroauxin IAA, 2.0mg/L and the agar of 3.4g/L of 6-benzyladenine 6-BA, 0.5mg/L of 3.0mg/L in MS propagating culture medium, the pH value of medium is 5.8, and adventitious buds proliferation coefficient is 17.9;
(4) Multiple Buds strong seedling culture: the test-tube plantlet Multiple Buds obtained in step (3) is placed in MS strong seedling culture base, at cultivation temperature 23-27 DEG C, intensity of illumination 1500lux, light application time is cultivate under the condition of 12-14 hour/day to obtain healthy and strong plant in 20 days, wherein contain heteroauxin IAA, 30g/L sucrose of 6-benzyladenine 6-BA, 1.0mg/L and the agar of 3.4g/L of 2.0mg/L in MS strong seedling culture base, the pH value of medium is 5.8, and the coefficient that Multiple Buds is bred again is 4.1;
(5) healthy and strong plant culture of rootage: the healthy and strong plant obtained in step (4) is placed in MS root media, at cultivation temperature 23-27 DEG C, intensity of illumination 1500lux, light application time is cultivate the whole plant obtaining for 35 days being with root under the condition of 12-14 hour/day, wherein contain root-inducing powder ABT, 30g/L sucrose of heteroauxin IAA, 0.3mg/L and the agar of 3.4g/L of 0.55mg/L in MS root media, the pH value of medium is 5.8, and rooting rate is 94.4%;
(6) hardening and transplanting: after step (5) obtains the whole plant being with root, bottle cap is opened in the indoor being 25 DEG C in room temperature, a small amount of running water is added in bottle, hardening 2-4 days, after surface horny is formed, seedling is taken out, clean root medium, be transplanted in husky bed immediately, grow one month in husky bed after, transplant land for growing field crops.
Embodiment 3
Another example of the quick breeding method for tissue culture of swordleaf dragon tree of the present invention, comprises the following steps:
(1) selection of explant and sterilization: get swordleaf dragon tree seed as explant, successively with 2% liquid detergent aqueous solution soaking 5min, wire tap water 15-30min, with the addition of 2-3 and drip 100 milliliter of 0.1% mercuric chloride sterilization 8-10min of Tween-20, aseptic water washing 3-5 time, finally remove surface moisture with sterilized filter paper, obtain explant, wherein sterile water is through autoclaved distilled water;
(2) seed germination obtains in vitro cuttings: be inoculated in MS medium by the explant that step (1) obtains, be 23-27 DEG C in cultivation temperature, intensity of illumination 1500lux, light application time is cultivate 30 days under the condition of 12-14 hour/day, in vitro cuttings is obtained after seed sprouting, wherein contain heteroauxin IAA, 30g/L sucrose of 6-benzyladenine 6-BA, 0.2mg/L and the agar of 3.4g/L of 1.0mg/L in MS medium, the pH value of medium is 5.8;
(3) test-tube plantlet Multiple Buds Fast-propagation is cultivated: the in vitro cuttings obtained in step (2) is placed in MS propagating culture medium, at cultivation temperature 23-27 DEG C, intensity of illumination 1500lux, light application time is cultivate under the condition of 8-10 hour/day to obtain test-tube plantlet Multiple Buds in 30 days, wherein contain kinetin KT, 30g/L sucrose of heteroauxin IAA, 1.05mg/L and the agar of 3.4g/L of 6-benzyladenine 6-BA, 0.3mg/L of 3.0mg/L in MS propagating culture medium, the pH value of medium is 5.8, and adventitious buds proliferation coefficient is 15.2;
(4) Multiple Buds strong seedling culture: the test-tube plantlet Multiple Buds obtained in step (3) is placed in MS strong seedling culture base, at cultivation temperature 23-27 DEG C, intensity of illumination 1500lux, light application time is cultivate under the condition of 12-14 hour/day to obtain healthy and strong plant in 20 days, wherein contain heteroauxin IAA, 30g/L sucrose of 6-benzyladenine 6-BA, 1.5mg/L and the agar of 3.4g/L of 3.0mg/L in MS strong seedling culture base, the pH value of medium is 5.8, and the coefficient that Multiple Buds is bred again is 5.0;
(5) healthy and strong plant culture of rootage: the healthy and strong plant obtained in step (4) is placed in MS root media, at cultivation temperature 23-27 DEG C, intensity of illumination 1500lux, light application time is cultivate the whole plant obtaining for 35 days being with root under the condition of 12-14 hour/day, wherein contain root-inducing powder ABT, 30g/L sucrose of heteroauxin IAA, 0.5mg/L and the agar of 3.4g/L of 1.0mg/L in MS root media, the pH value of medium is 5.8, and rooting rate is 98.2%;
(6) hardening and transplanting: after step (5) obtains the whole plant being with root, bottle cap is opened in the indoor being 25 DEG C in room temperature, a small amount of running water is added in bottle, hardening 2-4 days, after surface horny is formed, seedling is taken out, clean root medium, be transplanted in husky bed immediately, grow one month in husky bed after, transplant land for growing field crops.
Embodiment 4
Another example of the quick breeding method for tissue culture of swordleaf dragon tree of the present invention, comprises the following steps:
(1) selection of explant and sterilization: get swordleaf dragon tree seed as explant, successively with 2% liquid detergent aqueous solution soaking 5min, wire tap water 15-30min, with the addition of 2-3 and drip 100 milliliter of 0.1% mercuric chloride sterilization 8-10min of Tween-20, aseptic water washing 3-5 time, finally remove surface moisture with sterilized filter paper, obtain explant, wherein sterile water is through autoclaved distilled water;
(2) seed germination obtains in vitro cuttings: be inoculated in MS medium by the explant that step (1) obtains, be 23-27 DEG C in cultivation temperature, intensity of illumination 1500lux, light application time is cultivate 30 days under the condition of 12-14 hour/day, in vitro cuttings is obtained after seed sprouting, wherein contain heteroauxin IAA, 30g/L sucrose of 6-benzyladenine 6-BA, 0.2mg/L and the agar of 3.4g/L of 1.0mg/L in MS medium, the pH value of medium is 5.8;
(3) test-tube plantlet Multiple Buds Fast-propagation is cultivated: the in vitro cuttings obtained in step (2) is placed in MS propagating culture medium, at cultivation temperature 23-27 DEG C, intensity of illumination 1500lux, light application time is cultivate under the condition of 8-10 hour/day to obtain test-tube plantlet Multiple Buds in 30 days, wherein contain in MS propagating culture medium, kinetin KT, 30g/L sucrose of heteroauxin IAA, 0.1mg/L of 6-benzyladenine 6-BA, 0.1mg/L of 5.0mg/L and the agar of 3.4g/L, the pH value of medium is 5.8, and adventitious buds proliferation coefficient is 14.3;
(4) Multiple Buds strong seedling culture: the test-tube plantlet Multiple Buds obtained in step (3) is placed in MS strong seedling culture base, at cultivation temperature 23-27 DEG C, intensity of illumination 1500lux, light application time is cultivate under the condition of 12-14 hour/day to obtain healthy and strong plant in 20 days, wherein contain heteroauxin IAA, 30g/L sucrose of 6-benzyladenine 6-BA, 1.5mg/L and the agar of 3.4g/L of 1.0mg/L in MS strong seedling culture base, the pH value of medium is 5.8, and the coefficient that Multiple Buds is bred again is 3.8;
(5) healthy and strong plant culture of rootage: the healthy and strong plant obtained in step (4) is placed in MS root media, at cultivation temperature 23-27 DEG C, intensity of illumination 1500lux, light application time is cultivate the whole plant obtaining for 35 days being with root under the condition of 12-14 hour/day, wherein contain root-inducing powder ABT, 30g/L sucrose of heteroauxin IAA, 0.1mg/L and the agar of 3.4g/L of 1.0mg/L in MS root media, the pH value of medium is 5.8, and rooting rate is 96.7%;
(6) hardening and transplanting: after step (5) obtains the whole plant being with root, bottle cap is opened in the indoor being 25 DEG C in room temperature, a small amount of running water is added in bottle, hardening 2-4 days, after surface horny is formed, seedling is taken out, clean root medium, be transplanted in husky bed immediately, grow one month in husky bed after, transplant land for growing field crops.
Claims (1)
1. a quick breeding method for tissue culture for swordleaf dragon tree, is characterized in that: the method comprises the following steps:
(1) selection of explant and sterilization: get swordleaf dragon tree seed as explant, successively with 2% liquid detergent aqueous solution soaking 5min, wire tap water 15-30min, with the addition of 2-3 and drip 100 milliliter of 0.1% mercuric chloride sterilization 8-10min of Tween-20, aseptic water washing 3-5 time, finally remove surface moisture with sterilized filter paper, obtain explant, wherein sterile water is through autoclaved distilled water;
(2) seed germination obtains in vitro cuttings: be inoculated in MS medium by the explant that step (1) obtains, be 23-27 DEG C in cultivation temperature, intensity of illumination 1500lux, light application time is cultivate 30 days under the condition of 12-14 hour/day, in vitro cuttings is obtained after seed sprouting, wherein MS medium is minimal medium, add heteroauxin IAA, 30g/L sucrose of 6-benzyladenine 6-BA, 0.2mg/L of 1.0mg/L and the agar of 3.4g/L in medium, the pH value of medium is 5.8;
(3) test-tube plantlet Multiple Buds Fast-propagation is cultivated: the in vitro cuttings obtained in step (2) is placed in MS propagating culture medium, at cultivation temperature 23-27 DEG C, intensity of illumination 1500lux, light application time is cultivate under the condition of 8-10 hour/day to obtain test-tube plantlet Multiple Buds in 30 days, wherein MS propagating culture medium with MS medium for minimal medium, the 6-benzyladenine 6-BA of 1.0-5.0mg/L is added in medium, the heteroauxin IAA of 0.1-0.5mg/L, the kinetin KT of 0.1-2.0mg/L, the agar of 30g/L sucrose and 3.4g/L, the pH value of medium is 5.8,
(4) Multiple Buds strong seedling culture: the test-tube plantlet Multiple Buds obtained in step (3) is placed in MS strong seedling culture base, at cultivation temperature 23-27 DEG C, intensity of illumination 1500lux, light application time is cultivate under the condition of 12-14 hour/day to obtain healthy and strong plant in 20 days, wherein in MS strong seedling culture base with MS medium for minimal medium, add heteroauxin IAA, 30g/L sucrose of 6-benzyladenine 6-BA, 0.5-1.5mg/L of 1.0-3.0mg/L and the agar of 3.4g/L in medium, the pH value of medium is 5.8;
(5) healthy and strong plant culture of rootage: the healthy and strong plant obtained in step (4) is placed in MS root media, at cultivation temperature 23-27 DEG C, intensity of illumination 1500lux, light application time is cultivate the whole plant obtaining for 35 days being with root under the condition of 12-14 hour/day, wherein MS root media with MS medium for minimal medium, add root-inducing powder ABT, 30g/L sucrose of heteroauxin IAA, 0.1-0.5mg/L of 0.1-1.0mg/L and the agar of 3.4g/L in medium, the pH value of medium is 5.8;
(6) hardening and transplanting: after step (5) obtains the whole plant being with root, bottle cap is opened in the indoor being 25 DEG C in room temperature, a small amount of running water is added in bottle, hardening 2-4 days, after surface horny is formed, seedling is taken out, clean root medium, be transplanted in husky bed immediately, grow one month in husky bed after, transplant land for growing field crops.
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| CN105340736B (en) * | 2015-11-09 | 2017-12-19 | 广西壮族自治区药用植物园 | A kind of method of swordleaf dragon tree Lax callus induction |
| CN105532452B (en) * | 2015-12-09 | 2017-10-24 | 广西壮族自治区药用植物园 | A kind of mezereon Seed inducement and rapid propagation method |
| CN107242137B (en) * | 2017-08-04 | 2019-04-19 | 黄小燕 | The quick breeding method for tissue culture of cardiospermum halicacabum |
| CN108112476B (en) * | 2017-12-19 | 2019-01-01 | 佛山市三水阳特园艺有限公司 | A kind of Jin Xin Yemen iron quick breeding method for tissue culture |
| CN108391591B (en) * | 2018-01-26 | 2021-07-06 | 长江大学 | Tissue culture and rapid propagation method for tabebuia flavedo |
| CN108901858B (en) * | 2018-09-28 | 2020-02-14 | 恩施土家族苗族自治州农业科学院 | Rapid propagation method for selenium-rich protospecies of zingiber officinale roscoe |
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