CN103404432B - The tissue culture and rapid proliferation method of poinsettia - Google Patents
The tissue culture and rapid proliferation method of poinsettia Download PDFInfo
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Abstract
The present invention relates to a kind of tissue culture and rapid proliferation method of poinsettia, belong to field of plant tissue culture technique.It is characterized in that, utilize terminal bud or the lateral bud of potted plant poinsettia plantlet, aseptic culture medium is rapidly formed and breeds seedling in a large number, be transplanted in outdoor small flower after seedling root induction, survival rate more than 89%, grow 8 ~ 9 months, can be used as commodity selling.Poinsettia is produced and upper mainly to be bred by cuttage, but contains abundant milk because of branch, and cuttage survival rate is low; Tissue-culturing rapid propagation on the taking root of seedling difficult comparatively, rooting rate is low.The present invention is compared with traditional cottage propagation, and efficiency is high, and potted plant shapeliness, growing way are consistent, commodity value is high; The proliferation times of plantlet in vitro is 8 ~ 10 times, and propagation seedling rootage duration is 18 ~ 23 days, and rooting rate is 80 ~ 85%, substantially overcomes the problem that seedling is difficult to take root.The combination of seedling hormone in medium is simple, and cost is low, is convenient to promote.
Description
Technical field
The present invention relates to a kind of tissue cultures and method for quickly breeding of ornamental flower poinsettia, belong to biological technical field, specifically belong to Plant Tissue Breeding category.
Background technology
Poinsettia (
euphorbiapulcherrima), also claiming poinsettia, is Euphorbiaceae euphorbia, evergreen or half evergreen shrubs, and when blooming, leaf is vermilion, very beautiful, main for viewing and admiring.It originates in Central America Mexico one band, and all there is cultivation existing China various places.Its ornamental value and economic worth are all higher.
At New Year's Day, Spring Festival, most of flowers and trees are also in the dormant stage, and poinsettia then wears greenery, and strain jacking is with crimson gorgeous petal-shaped bract, and the large look of shape is fresh, very noticeable, are the excellent flowers in red-letter day in winter, so be " Christmas flower " also known as it.
Poinsettia is rare sets seeds or substantially without the ability of setting seeds, production is bred by cuttage, but contains abundant milk due to branch, and cuttage survival rate is low, and some improved seeds branches are few in addition, and it is limited to introduce a fine variety initial stage reproductive number, is difficult to meet consumer demand.
At present, the research of the tissue-culturing rapid propagation of domestic poinsettia is few, not yet forms ripe cultivating system, especially also acquires a certain degree of difficulty on the taking root of seedling.The present invention for material, by constantly groping and improveing, obtains the tissue-culturing rapid propagation system of a set of maturation with potted plant poinsettia plantlet, for the large-scale production of poinsettia seedling provides technical reserve.
Summary of the invention
The object of the invention is to, provide a kind of cycle short, efficiency is high, and production cost is low, the method for energy scale Fast-propagation commodity poinsettia potted flower.
Technical scheme of the present invention, the steps include:
(1) explant sterilization process: choose without damage by disease and insect, robust growth, clean potted plant poinsettia plantlet; Use sterilizing scissors, the clip plant top tender terminal bud of children and 2 ~ 5 lateral buds, add a small amount of liquid detergent, with tap water 3 ~ 5min; Put into sterile beaker, on super-clean bench, first embathe 4 ~ 5 times with sterile water, each 30 ~ 60S; Use 75% alcohol-pickled 50 ~ 60S again, tipping alcohol; Then 0.1%HgCl is used
2solution sterilization 8 ~ 10min, tipping waste liquid; Finally use aseptic water washing 5 ~ 7 times, by material transfer in the sterile petri dish being placed with blotting paper, suck dry moisture;
(2) induction of aseptic seedling and Initial culture: by above-mentioned sterilization treatment, the stem explants of band terminal bud or lateral bud is seeded in Initial culture base (MS+6-BA1.0 ~ 1.5mg/L+NAA0.1 ~ 0.15mg/L+0.6% agar+3% sucrose, pH value is 5.8 ~ 6.0) on, cultivation temperature is 22 ~ 24 DEG C, illuminance is 2000 ~ 2200Lx, and light application time is 12h/d; 24 ~ 27 days, cutting shoots grew up to the seedling of high 2.5 ~ 4.3cm, seedling leaf dark green, robust growth;
(3) propagation of seedling and squamous subculture: by the seedling of above-mentioned Initial culture, be cut into the segment of 2 ~ 4 band buds, be seeded in subculture medium (MS+6-BA1.0 ~ 1.5mg/L+KT0.5 ~ 1.0mg/L+IAA0.4 ~ 0.6mg/L+0.6% agar+3% sucrose, pH value is 5.8 ~ 6.0) on, cultivation temperature is 22 ~ 24 DEG C, illuminance is 2000 ~ 2200Lx, and light application time is 12h/d; 20 ~ 25 days, segment grew up to again high 3.3 ~ 5.0cm, leaf look dark green seedling of growing thickly, and proliferation times is 8 ~ 10 times;
(4) culture of rootage of seedling: choose the seedling through squamous subculture, growing way stalwartness, intercept from base portion near joint position, be linked into root media (1/2MS+IAA0.5 ~ 0.8mg/L+IBA0.5 ~ 0.8mg/L+0.5% agar+2% sucrose, pH value is 5.8 ~ 6.0) in, cultivation temperature is 22 ~ 24 DEG C, illuminance is 2000 ~ 2200Lx, and light application time is 12h/d; 18 ~ 23 days, the root that seedling base portion has 4 ~ 6 1.0 ~ 2.5cm generated, and rooting rate is 80 ~ 85%;
(5) transplanting of seedling: seedling of taking root grows in bottle that cane is sturdy, leaf look dark green, high 4.5 ~ 6.0cm time, carry out flowerpot transplanting, first bottle removed lid, place 2 ~ 3 days in culturing room, keep the skin wet at any time according to the growing state of seedling therebetween; Through KMnO
4the diameter of sterilization is in the small flower of 16cm, puts into the detritus soil through high-temperature sterilization, moves in flowerpot by bottle seedling, water sufficient water, places 3 ~ 4 days in culturing room, moves into greenhouse; Within 2 ~ 3 months, turn basin once.
The invention has the beneficial effects as follows: utilize the poinsettia plant that 3-4 strain is potted plant, by tissue culture technique, produce the commercial poinsettia plant of thousands of basin fast, its feature is as follows:
(1) efficiency is high, and the cycle is shorter, and potted plant shapeliness, growing way are strong, commodity value is high, in table 1;
Table 1 the inventive method compares with conventional method
| method | quantity | feature |
| tradition cuttage or division propagation | one basin can plant division 3-6 basin | from the branch that maternal plant is sheared, base portion conduit is easily blocked by milk, and Water Transportation is obstructed, and survival rate is low, difficulty of taking root, plant growing way are poor, from plant division to commodity selling, need 10 ~ 12 months. |
| the tissue-culturing rapid propagation that the present invention relates to | one basin can obtain several thousand even several ten thousand basins | the plantlet in vitro of having taken root because root system is many, root is sturdy, the ability that absorbs moisture and nutrition is stronger, growth is fast, plant forming, commodity value are high, from being transplanted into flowerpot to commodity selling needs 8 ~ 9 months. |
(2) the short rooting rate height the present invention of the cycle of plantlet in vitro is from harvesting explant bud, and to the seedling that acquisition is taken root, whole process approximately needs about 80 days, and the cycle is shorter, and overcomes the problem that seedling is difficult to take root, and rooting rate is higher;
(3) in the lower culture medium prescription of the present invention of production cost, employ several routine growth hormone (6-BA, KT, NAA, IAA, IBA), hormone combinations is simple, and cost is not high.
Embodiment
Following examples of implementation further illustrate of the present invention, is not limitation of the present invention.
example one:
Choose without damage by disease and insect, robust growth, clean potted plant poinsettia plantlet; Use sterilizing scissors, the clip plant top tender terminal bud of children and 2 lateral buds, add a small amount of liquid detergent, use tap water 5min; Put into sterile beaker, on super-clean bench, first embathe 4 times with sterile water, each 30S; Use 75% alcohol-pickled 60S again, tipping alcohol; Then 0.1%HgCl is used
2solution sterilization 10min, tipping waste liquid; Finally use aseptic water washing 7 times, by material transfer in the sterile petri dish being placed with blotting paper, suck dry moisture;
By above-mentioned sterilization treatment, band terminal bud or the stem explants of lateral bud be seeded in Initial culture base (MS+6-BA1.0mg/L+NAA0.1mg/L+0.6% agar+3% sucrose, pH value is 5.8) on, cultivation temperature is 22 DEG C, and illuminance is 2000Lx, and light application time is 12h/d; 25 days, cutting shoots grew up to the seedling of high 3.0cm, seedling leaf dark green, robust growth;
By the seedling of above-mentioned Initial culture, be cut into the segment of 3 band buds, be seeded in subculture medium (MS+6-BA1.0mg/L+KT0.5mg/L+IAA0.5mg/L+0.6% agar+3% sucrose, pH value is 5.8) on, cultivation temperature is 22 DEG C, and illuminance is 2000Lx, and light application time is 12h/d; 22 days, segment grew up to again high 4.0cm, leaf look dark green seedling of growing thickly, and proliferation times is 9 times;
Choose the seedling through squamous subculture, growing way stalwartness, intercept near joint position from base portion, be linked into root media (1/2MS+IAA0.5mg/L+IBA0.5mg/L+0.5% agar+2% sucrose, pH value is 5.8) in, cultivation temperature is 22 DEG C, and illuminance is 2000Lx, and light application time is 12h/d; 20 days, the root that seedling base portion has 6 1.0 ~ 2.5cm generated, and rooting rate is 80%;
Seedling of taking root grows in bottle that cane is sturdy, leaf look dark green, high 4.5cm time, carry out flowerpot transplanting, first bottle removed lid, place 2 days in culturing room, keep the skin wet at any time according to the growing state of seedling therebetween; Through KMnO
4the diameter of sterilization is in the small flower of 16cm, puts into the detritus soil through high-temperature sterilization, moves in flowerpot by bottle seedling, water sufficient water, places 3 days in culturing room, moves into greenhouse; Within 2 months, turn basin once.
example two:
Choose without damage by disease and insect, robust growth, clean potted plant poinsettia plantlet; Use sterilizing scissors, the clip plant top tender terminal bud of children and 3 lateral buds, add a small amount of liquid detergent, use tap water 3min; Put into sterile beaker, on super-clean bench, first soak 5 times with sterile water, each 60S; Use 75% alcohol-pickled 50S again, tipping alcohol; Then 0.1%HgCl is used
2solution sterilization 9min, tipping waste liquid; Finally use aseptic water washing 5 times, by material transfer in the sterile petri dish being placed with blotting paper, suck dry moisture;
By above-mentioned sterilization treatment, band terminal bud or the stem explants of lateral bud be seeded in Initial culture base (MS+6-BA1.5mg/L+NAA0.15mg/L+0.6% agar+3% sucrose, pH value is 6.0) on, cultivation temperature is 24 DEG C, and illuminance is 2200Lx, and light application time is 12h/d; 27 days, cutting shoots grew up to the seedling of high 4.3cm, seedling leaf dark green, robust growth;
By the seedling of above-mentioned Initial culture, be cut into the segment of 4 band buds, be seeded in subculture medium (MS+6-BA1.5mg/L+KT1.0mg/L+IAA0.4mg/L+0.6% agar+3% sucrose, pH value is 6.0) on, cultivation temperature is 24 DEG C, and illuminance is 2200Lx, and light application time is 12h/d; 25 days, segment grew up to again high 5.0cm, leaf look dark green seedling of growing thickly, and proliferation times is 8 times;
Choose the seedling through squamous subculture, growing way stalwartness, intercept near joint position from base portion, be linked into root media (1/2MS+IAA0.8mg/L+IBA0.6mg/L+0.5% agar+2% sucrose, pH value is 6.0) in, cultivation temperature is 24 DEG C, and illuminance is 2200Lx, and light application time is 12h/d; 23 days, the root that seedling base portion has 5 1.0 ~ 2.5cm generated, and rooting rate is 85%;
Seedling of taking root grows in bottle that cane is sturdy, leaf look dark green, high 6.0cm time, carry out flowerpot transplanting, first bottle removed lid, place 3 days in culturing room, keep the skin wet at any time according to the growing state of seedling therebetween; Through KMnO
4the diameter of sterilization is in the small flower of 16cm, puts into the detritus soil through high-temperature sterilization, moves in flowerpot by bottle seedling, water sufficient water, places 4 days in culturing room, moves into greenhouse; Within 3 months, turn basin once.
example three:
Choose without damage by disease and insect, robust growth, clean potted plant poinsettia plantlet; Use sterilizing scissors, the clip plant top tender terminal bud of children and 5 lateral buds, add a small amount of liquid detergent, use tap water 4min; Put into sterile beaker, on super-clean bench, first soak 4 times with sterile water, each 45S; Use 75% alcohol-pickled 55S again, tipping alcohol; Then 0.1%HgCl is used
2solution sterilization 8min, tipping waste liquid; Finally use aseptic water washing 6 times, by material transfer in the sterile petri dish being placed with blotting paper, suck dry moisture;
By above-mentioned sterilization treatment, band terminal bud or the stem explants of lateral bud be seeded in Initial culture base (MS+6-BA1.2mg/L+NAA0.13mg/L+0.6% agar+3% sucrose, pH value is 5.8) on, cultivation temperature is 23 DEG C, and illuminance is 2000Lx, and light application time is 12h/d; 24 days, cutting shoots grew up to the seedling of high 2.5cm, seedling leaf dark green, robust growth;
By the seedling of above-mentioned Initial culture, be cut into the segment of 2 band buds, be seeded in subculture medium (MS+6-BA1.2mg/L+KT0.8mg/L+IAA0.6mg/L+0.6% agar+3% sucrose, pH value is 5.8) on, cultivation temperature is 23 DEG C, and illuminance is 2000Lx, and light application time is 12h/d; 20 days, segment grew up to again high 3.3cm, leaf look dark green seedling of growing thickly, and proliferation times is 10 times;
Choose the seedling through squamous subculture, growing way stalwartness, intercept near joint position from base portion, be linked into root media (1/2MS+IAA0.6mg/L+IBA0.8mg/L+0.5% agar+2% sucrose, pH value is 5.8) in, cultivation temperature is 23 DEG C, and illuminance is 2000Lx, and light application time is 12h/d; 18 days, the root that seedling base portion has 4 1.0 ~ 2.5cm generated, and rooting rate is 83%;
The transplanting of seedling: seedling of taking root grows in bottle that cane is sturdy, leaf look dark green, high 5.5cm time, carry out flowerpot transplanting, first bottle removed lid, place 3 days in culturing room, keep the skin wet at any time according to the growing state of seedling therebetween; Through KMnO
4the diameter of sterilization is in the small flower of 16cm, puts into the detritus soil through high-temperature sterilization, moves in flowerpot by bottle seedling, water sufficient water, places 3 days in culturing room, moves into greenhouse; 2 first quarter moons turn basin once.
example four:
Choose without damage by disease and insect, robust growth, clean potted plant poinsettia plantlet; Use sterilizing scissors, the clip plant top tender terminal bud of children and 3 lateral buds, add a small amount of liquid detergent, use tap water 5min; Put into sterile beaker, on super-clean bench, first soak 5 times with sterile water, each 60S; Use 75% alcohol-pickled 60S again, tipping alcohol; Then 0.1%HgCl is used
2solution sterilization 10min, tipping waste liquid; Finally use aseptic water washing 6 times, by material transfer in the sterile petri dish being placed with blotting paper, suck dry moisture;
By above-mentioned sterilization treatment, band terminal bud or the stem explants of lateral bud be seeded in Initial culture base (MS+6-BA1.0mg/L+NAA0.15mg/L+0.6% agar+3% sucrose, pH value is 5.8) on, cultivation temperature is 23 DEG C, and illuminance is 2200Lx, and light application time is 12h/d; 26 days, cutting shoots grew up to the seedling of high 3.8cm, seedling leaf dark green, robust growth;
By the seedling of above-mentioned Initial culture, be cut into the segment of 3 band buds, be seeded in subculture medium (MS+6-BA1.5mg/L+KT0.5mg/L+IAA0.5mg/L+0.6% agar+3% sucrose, pH value is 5.8) on, cultivation temperature is 23 DEG C, and illuminance is 2200Lx, and light application time is 12h/d; 23 days, segment grew up to again high 4.5cm, leaf look dark green seedling of growing thickly, and proliferation times is 9 times;
Choose the seedling through squamous subculture, growing way stalwartness, intercept near joint position from base portion, be linked into root media (1/2MS+IAA0.7mg/L+IBA0.7mg/L+0.5% agar+2% sucrose, pH value is 5.8) in, cultivation temperature is 23 DEG C, and illuminance is 2200Lx, and light application time is 12h/d; 22 days, the root that seedling base portion has 6 1.0 ~ 2.5cm generated, and rooting rate is 85%;
The transplanting of seedling: seedling of taking root grows in bottle that cane is sturdy, leaf look dark green, high 5.0cm time, carry out flowerpot transplanting, first bottle removed lid, place 3 days in culturing room, keep the skin wet at any time according to the growing state of seedling therebetween; Through KMnO
4the diameter of sterilization is in the small flower of 16cm, puts into the detritus soil through high-temperature sterilization, moves in flowerpot by bottle seedling, water sufficient water, places 3 days in culturing room, moves into greenhouse; Within 3 months, turn basin once.
Claims (1)
1. a tissue culture and rapid proliferation method for poinsettia, is characterized in that, comprise the following steps:
(1) explant sterilization process: choose without damage by disease and insect, robust growth, clean potted plant poinsettia plantlet; Use sterilizing scissors, the clip plant top tender terminal bud of children and 2 ~ 5 lateral buds, add a small amount of liquid detergent, with tap water 3 ~ 5min; Put into sterile beaker, on super-clean bench, first embathe 4 ~ 5 times with sterile water, each 30 ~ 60S; Use 75% alcohol-pickled 50 ~ 60S again, tipping alcohol; Then 0.1%HgCl is used
2solution sterilization 8 ~ 10min, tipping waste liquid; Finally use aseptic water washing 5 ~ 7 times, by material transfer in the sterile petri dish being placed with blotting paper, suck dry moisture;
(2) induction of aseptic seedling and Initial culture: by above-mentioned sterilization treatment, the stem explants of band terminal bud or lateral bud is seeded on Initial culture base, described medium is: MS+6-benzylaminopurine (6-BA) 1.0 ~ 1.5mg/L+ methyl α-naphthyl acetate (NAA) 0.1 ~ 0.15mg/L+0.6% agar+3% sucrose, adjustment medium pH value is 5.8 ~ 6.0, cultivation temperature is 22 ~ 24 DEG C, illuminance is 2000 ~ 2200Lx, and light application time is 12h/d; 24 ~ 27 days, cutting shoots grew up to the seedling of high 2.5 ~ 4.3cm, seedling leaf dark green, robust growth;
(3) propagation of seedling and squamous subculture: by the seedling of above-mentioned Initial culture, be cut into the segment of 2 ~ 4 band buds, be seeded on subculture medium, described medium is: MS+6-BA1.0 ~ 1.5mg/L+ kinetin (KT) 0.5 ~ 1.0mg/L+ heteroauxin (IAA) 0.4 ~ 0.6mg/L+0.6% agar+3% sucrose, adjustment medium pH value is 5.8 ~ 6.0, cultivation temperature is 22 ~ 24 DEG C, and illuminance is 2000 ~ 2200Lx, and light application time is 12h/d; 20 ~ 25 days, segment grew up to high 3.3 ~ 5.0cm, leaf look dark green seedling of growing thickly, and proliferation times is 8 ~ 10 times;
(4) culture of rootage of seedling: choose the seedling through squamous subculture, growing way stalwartness, intercept from base portion near joint position, be linked in root media, described medium is: 1/2MS+IAA0.5 ~ 0.8mg/L+IBA0.5 ~ 0.8mg/L+0.5% agar+2% sucrose, adjustment medium pH value is 5.8 ~ 6.0, cultivation temperature is 22 ~ 24 DEG C, and illuminance is 2000 ~ 2200Lx, and light application time is 12h/d; 18 ~ 23 days, the root that seedling base portion has 4 ~ 6 1.0 ~ 2.5cm generated, and rooting rate is 80 ~ 85%;
(5) transplanting of seedling: seedling of taking root grows in bottle that cane is sturdy, leaf look dark green, high 4.5 ~ 6.0cm time, carry out following flowerpot transplanting: first bottle is removed lid, place 2 ~ 3 days in culturing room, keep the skin wet at any time according to the growing state of seedling therebetween; Through KMnO
4the diameter of sterilization is in the small flower of 16cm, puts into the detritus soil through high-temperature sterilization, moves in flowerpot by bottle seedling, water sufficient water, places 3 ~ 4 days in culturing room, moves into greenhouse; Within 2 ~ 3 months, turn basin once, grow 8 ~ 9 months; The little shoot survival percent of transplanting through this method is more than 89%.
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| CN107616090A (en) * | 2017-11-14 | 2018-01-23 | 佴辉 | Poinsettia breeding of new variety method and its scale rapid propagation method |
| CN108293879B (en) * | 2018-04-20 | 2021-06-01 | 西北大学 | Rapid propagation method of euphorbia kansui |
| CN109618925B (en) * | 2018-11-07 | 2021-10-01 | 中南林业科技大学 | A method suitable for rapid propagation of various shrubs and herbaceous flower plants |
| CN112715357B (en) * | 2020-12-17 | 2022-01-04 | 广州甘蔗糖业研究所湛江甘蔗研究中心 | A Tissue Culture Rapid Propagation Method of Prunus chinensis suitable for factory production |
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| US4634674A (en) * | 1983-03-25 | 1987-01-06 | Atlantic Richfield Company | Plant regeneration from protoplasts |
| CN102845312A (en) * | 2012-10-12 | 2013-01-02 | 江苏省农业科学院 | Method for obtaining poinsettia filial generation through utilizing young embryo rescue |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4634674A (en) * | 1983-03-25 | 1987-01-06 | Atlantic Richfield Company | Plant regeneration from protoplasts |
| CN102845312A (en) * | 2012-10-12 | 2013-01-02 | 江苏省农业科学院 | Method for obtaining poinsettia filial generation through utilizing young embryo rescue |
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| Title |
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| 一品红柠檬雪茎段组织培养快繁技术;陈璐;《江苏农业科学》;20111215;第39卷(第6期);第50页摘要 * |
| 一品红组培快繁技术的初步研究;曹帮华;《山东林业科技》;19991022(第5期);第7页摘要，第1.1-1.4节，第8页第1.5节 * |
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