CN102754598B - Method for inducing test-tube bulblet of lily - Google Patents
Method for inducing test-tube bulblet of lily Download PDFInfo
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- CN102754598B CN102754598B CN 201210237261 CN201210237261A CN102754598B CN 102754598 B CN102754598 B CN 102754598B CN 201210237261 CN201210237261 CN 201210237261 CN 201210237261 A CN201210237261 A CN 201210237261A CN 102754598 B CN102754598 B CN 102754598B
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Abstract
The invention discloses a method for inducing a test-tube bulblet of lily. The method disclosed by the invention comprises the following steps of: inoculating the lily bulb as an explant into a differential culture medium; then firstly inducing to generate cluster seedlings, and then carrying out low temperature treatment on the cluster seedlings; inoculating the cluster seedlings into a bulb induction culture medium; inducing the test-tube bulblet; carrying out low temperature treatment at the temperature of 10 DEG C for 14 days; and culturing in a constant temperature culture room, thus being beneficial to expansion of the test-tube bulblet. A robust lily bulblet is induced in a test tube, thus growth time of tissue culture seedling can be beneficially shortened and production cost can be reduced.
Description
Technical field
The present invention relates to the fast breeding method of a kind of lily, be specifically related to a kind of abductive approach of bulblet of lilium.
Background technology
For a long time, lily breeds in modes such as traditional bulb separation, branch bulbil, scale cuttage, scale embeddings always, has deficiencies such as reproduction coefficient is little, kind sexual involution, virus accumulation.
Summary of the invention
At the deficiencies in the prior art, purpose of the present invention will provide a kind of shortening group training seedling production time exactly, reduces cost, and sloughs plant body inner virus effectively, prevents kind of a sexual involution, cultivates the abductive approach of the low bulblet of lilium of cost.
The object of the present invention is achieved like this: a kind of abductive approach of bulblet of lilium may further comprise the steps:
1) scale low temperature treatment: select robust growth lily bulb ball, clean back refrigeration 7-10 days;
2) explant sterilization: will peel off the 1-2 layer scale of outside through the lily bulb ball of low temperature storage, choosing inner scale is explant, with explant clean the back on superclean bench earlier with alcohol-pickled, sterilize with chlorization mercury liquid again, and with behind the aseptic water washing scale being placed on the aseptic filter paper, dice is inoculated in to start on the differential medium and cultivates;
3) start to cultivate: it is that the constant temperature culture of 25 ± 1 ℃ of temperature, humidity 75 ± 2%, illumination 14h, intensity of illumination 1800-2000lx is indoor that the differential medium of the explant that inoculation is had places environmental condition, cultivate after 15 days and produce white bulk callus from lily incision and the dedifferentiation of belly edge, continue the cultivation by 20 days, break up the bud of growing thickly on the white bulk callus again;
4) bulb is induced cultivation: when the bud length of growing thickly arrives 3-5cm, the bud of will growing thickly carries out strong seedling culture after being divided into little simple bud, healthy and strong seedling placed in 10 ℃ of illumination boxs handle 14d, be transferred to again in the bulb inducing culture, there is the bulb inducing culture of healthy and strong seedling to place the indoor bulblet of lilium that carries out in vitro of constant temperature culture to induce switching, cultivated by 45 days and induce bulb.
Step 2) and 3) described differential medium prescription is:
Ms+6-BA1.5mg/L+NAA0.5mg/L+IBA0.1mg/L+ sucrose 30g/L+ agar powder 5g/L.
The prescription of the bulb inducing culture described in the step 4) is:
Ms+6-BA0.5 mg/L+NAA0.1 mg/L+sucrose 30 g/L+ agar powders 5 mg/L.
The refrigerated storage temperature of refrigerator was 5 ℃ when step 1) was selected explant bulb ball low temperature treatment for use, time 7-10 days.
In the step 4) switching is had the bulb inducing culture of healthy and strong seedling to place when constant temperature culture is indoor carries out that in vitro bulblet of lilium is induced, the indoor temperature of constant temperature culture is 10 ± 1 ℃, humidity 75 ± 2%, illumination 14h, intensity of illumination 1800-2000lx..
The abductive approach of bulblet of lilium provided by the invention has following beneficial effect:
1, lily to use the lily bulb through low temperature storage be explant, be seeded in medium and start and induce the seedling of growing thickly; To grow thickly again seedling with 10 ℃ of low temperature treatment 14d after, be transferred to the bulb inducing culture and induce the test tube clove, produce the bud of growing thickly than directly inducing, in vitro do not form healthy and strong clove transplant survival height, cost is low.
2, lily is directly induced clove from test tube, can be with the direct acclimatization and transplants of clove, the training seedling production time of shortening group greatly, reduce cost.
3, lily is cultivated to induce by continuous test tube subculture and produces clove, can slough plant body inner virus effectively, prevents kind of a sexual involution.
Embodiment
Differential medium preparation: be minimal medium with Ms, add sucrose 30g/L and agar powder 5g/L in the medium, the hormone combinations of the interpolation in the medium is: 6-BA1.5mg/L+NAA0.5mg/L+IBA0.1mg/L;
Bulb inducing culture preparation: be minimal medium with Ms, add sucrose 30g/L and agar powder 5g/L in the medium, the hormone combinations of the interpolation in the medium is: 6-BA0.5 mg/L+NAA0.1 mg/L.
Wherein: 6-BA is that 6-benzyladenine, NAA are that naa, IBA are indolebutyric acids.
Induce process instance:
1) scale low temperature treatment: select robust growth lily bulb ball, clean and be placed on 5 ℃ of refrigerators and refrigerate about 7-10 days;
2) explant sterilization: will peel off the 1-2 layer scale of outside through the bulb of low temperature storage, choosing inner scale is explant, place 5% washing powder solution to soak 10min and cleaning explant, place flowing water flushing 1h again, aseptic water washing earlier with 75% alcohol-pickled 30s, through 0.1% liter of chlorization mercury liquid (1g/L of unit) sterilization 15min, is used 5 times again in the flushing back on superclean bench, explant scale after the flushing places on the aseptic filter paper, is cut into 1cm
2Square is inoculated in to start on the differential medium and cultivates;
3) start to cultivate: having the differential medium of explant to place environmental condition inoculation is that the constant temperature culture of 25 ± 1 ℃ of temperature, humidity 75 ± 2%, illumination 14h, intensity of illumination 1800-2000lx is indoor, cultivate and produce white callus from lily incision and the dedifferentiation of belly edge after 15 days, continue by breaking up the bud of growing thickly again on 20 days the cultivation white bulk callus;
4) bulb is induced cultivation: when the bud length of growing thickly arrives 3-5cm, carry out strong seedling culture after being divided into little simple bud, healthy and strong seedling placed in 10 ℃ of illumination boxs handle 14d, after being transferred to the bulb inducing culture again, place and carry out in vitro clove into the constant temperature culture chamber and induce, the indoor temperature of constant temperature culture is 10 ± 1 ℃, humidity 75 ± 2%, illumination 14h, intensity of illumination 1800-2000lx..Cultivated the average fresh weight that induces bulb by 45 days and reach 1.4g/, the bulb color is creamy white, and the seedling acclimatization and transplants survival rate of growing thickly reaches more than 90% the seedling robust growth.
Claims (2)
1. the abductive approach of a bulblet of lilium is characterized in that: may further comprise the steps:
1) scale low temperature treatment: select robust growth lily bulb ball, clean back refrigeration 7-10 days;
2) explant sterilization: will peel off the 1-2 layer scale of outside through the lily bulb ball of low temperature storage, choosing inner scale is explant, with explant clean the back on superclean bench earlier with alcohol-pickled, sterilize with chlorization mercury liquid again, and with behind the aseptic water washing scale being placed on the aseptic filter paper, dice is inoculated in to start on the differential medium and cultivates;
3) start to cultivate: having the differential medium of explant to place environmental condition inoculation is that the constant temperature culture of 25 ± 1 ℃ of temperature, humidity 75 ± 2%, illumination 14h, intensity of illumination 1800-2000lx is indoor, cultivate after 15 days and produce white bulk callus from lily incision and the dedifferentiation of belly edge, continue the cultivation by 20 days, break up the bud of growing thickly on the white bulk callus again;
4) bulb is induced cultivation: when the bud length of growing thickly arrives 3-5cm, the bud of will growing thickly carries out strong seedling culture after being divided into little simple bud, healthy and strong seedling placed in 10 ℃ of illumination boxs handle 14d, be transferred to again in the bulb inducing culture, there is the bulb inducing culture of healthy and strong seedling to place the indoor bulblet of lilium that carries out in vitro of constant temperature culture to induce switching, cultivated by 45 days and induce bulb;
Step 2) and 3) described differential medium prescription is: MS+6-BA1.5mg/L+NAA0.5mg/L+IBA0.1mg/L+ sucrose 30g/L+ agar powder 5g/L;
The prescription of the bulb inducing culture described in the step 4) is: bulb inducing culture MS+6-BA0.5 mg/L+NAA0.1 mg/L+sucrose 30 g/L+agar powder 5 mg/L;
In the step 4) switching is had the bulb inducing culture of healthy and strong seedling to place when constant temperature culture is indoor carries out that in vitro bulblet of lilium is induced, the indoor temperature of constant temperature culture is 10 ± 1 ℃, humidity 75 ± 2%, illumination 14h, intensity of illumination 1800-2000lx.
2. the abductive approach of bulblet of lilium according to claim 1 is characterized in that: in the step 1) during lily bulb ball low temperature treatment refrigerated storage temperature of refrigerator be 5 ℃, time 7-10 days.
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Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103004588B (en) * | 2012-12-03 | 2014-09-24 | 中国计量学院 | Tissue culture and rapid propagation method of tulipa edulis |
| CN103314863B (en) * | 2013-07-16 | 2016-01-20 | 方倩 | A kind of method of efficient acquisition tulip callus |
| CN103385171B (en) * | 2013-07-16 | 2016-01-27 | 河南科技学院 | The breeding method of a kind of virus-free tulip seedling |
| CN103314864B (en) * | 2013-07-16 | 2015-05-06 | 李锋 | Method for obtaining detoxified seedling from bulb scales of tulip |
| CN103733994B (en) * | 2013-12-18 | 2016-01-06 | 浙江大学 | The method in a kind of oriental hybrid lily plantlet in vitro strong sprout |
| CN115500263B (en) * | 2022-10-11 | 2023-05-02 | 北京市农林科学院 | Method for rapidly inducing lily scales and expanding test tube bulblets |
| CN116391622B (en) * | 2023-04-19 | 2024-04-30 | 南京农业大学 | Lily tissue culture rapid propagation method and application thereof |
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| CN100374011C (en) * | 2003-11-07 | 2008-03-12 | 云南省农业科学院园艺作物研究所 | Method for tissue culture of lily flowers |
| CN100394845C (en) * | 2004-10-21 | 2008-06-18 | 云南省农业科学院花卉研究所 | In-bottle production method of detoxified small seed ball of east lily |
| CN100361570C (en) * | 2005-11-15 | 2008-01-16 | 中国热带农业科学院热带生物技术研究所 | Efficient in vifro culture method of fresh flower lily |
| CN100429306C (en) * | 2005-12-31 | 2008-10-29 | 南京大学 | Tissue culture medium and fast propagation method for Sorbone lily |
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